KRAS mutant protein inhibitors

ABSTRACT

The invention relates to a KRAS mutant protein inhibitor shown as formula (I), a composition containing the inhibitor and the use thereof.

TECHNICAL FIELD

The invention relates to a KRAS mutant protein inhibitors, a compositioncontaining the inhibitors and the use thereof.

BACKGROUND ART

RAS represents a population of 189 amino acid monomeric globularproteins (21 kDa molecular weight) that are associated with the plasmamembrane and bind to GDP or GTP, and RAS acts as a molecular switch.When the RAS contains bound GDP, it is in a stationary or closedposition and is “inactive”. When cells are exposed to certaingrowth-promoting stimuli, RAS is induced to exchange their bound GDP forGTP. In the case of binding to GTP, RAS is “opened” and is capable ofinteracting with other proteins (its “downstream targets”) andactivating the proteins. The RAS protein itself has an inherently lowability to hydrolyze GTP back to GDP, thereby turning itself into aclosed state. Closing RAS requires an exogenous protein called GTPaseactivating protein (GAP) that interacts with RAS and greatly acceleratesthe conversion of GTP to GDP. Any mutation in RAS that affects itsability to interact with GAP or convert GTP back to GDP will result inprolonged protein activation, and thus conduction to the cell to informits signaling of continued growth and division. Since these signalscause cell growth and division, over-activated RAS signaling canultimately lead to cancer.

Structurally, the RAS protein contains a G domain responsible for theenzymatic activity of RAS, guanine nucleotide binding and hydrolysis(GTPase reaction). It also contains a C-terminal extension called theCAAX cassette, which can be post-translationally modified andresponsible for targeting the protein to the membrane. The G domaincontains a phosphate binding ring (P-ring). The P-loop represents apocket of a binding nucleotide in a protein, and this is a rigid portionof a domain with conserved amino acid residues necessary for nucleotidebinding and hydrolysis (glycine 12 and lysine 16). The G domain alsocontains a so-called switch I region (residues 30-40) and a switch IIregion (residues 60-76), both of which are dynamic parts of the protein,since the dynamic portion is converted between stationary and loadedstates. The ability is often expressed as a “spring loaded” mechanism.The primary interaction is the hydrogen bond formed by threonine-35 andglycine-60 with the gamma-phosphate of GTP, which maintains the activeconformation of the switch 1 region and the switch 2 region,respectively. After hydrolysis of GTP and release of phosphate, the tworelax into an inactive GDP conformation.

The most notable members of the RAS subfamily are HRAS, KRAS and NRAS,which are primarily involved in many types of cancer. Mutation of any ofthe three major isoforms of the RAS gene (HRAS, NRAS or KRAS) is one ofthe most common events in human tumor formation. Approximately 30% ofall tumors in human tumors were found to carry some mutations in the RASgene. It is worth noting that KRAS mutations were detected in 25%-30% oftumors. In contrast, the rate of carcinogenic mutations in NRAS and HRASfamily members was much lower (8% and 3%, respectively). The most commonKRAS mutations were found at residues G12 and G13 in the P-loop as wellas at residue Q61.

G12C is a frequently occurring KRAS gene mutation (glycine-12 is mutatedto cysteine). This mutation has been found in about 13% of cancers,about 43% in lung cancer, and almost 100% in MYH-associated polyposis(familial colon cancer syndrome). However, targeting this gene withsmall molecules is a challenge.

Thus, despite advances in this field, there remains a need in the artfor improved compounds and methods for treating cancer, such as byinhibiting KRAS, HRAS or NRAS. The present invention fulfills this needand provides other related advantages.

SUMMARY OF INVENTION

In one aspect, provided herein is A compound of formula (I), astereoisomer thereof, an atropisomer thereof, a pharmaceuticallyacceptable salt thereof, a pharmaceutically acceptable salt of thestereoisomer thereof or a pharmaceutically acceptable salt of theatropisomer thereof:

Wherein:

R₁₁, R₁₂, R₁₃, R₁₄ or R₁₅ is independently selected from halogen,—C₁₋₆alkyl, —C₂₋₆alkenyl, —C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN,—OC₁₋₆alkyl, —SC₁₋₆alkyl, —NR₅R₆, —C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅,—C(═O)NR₅R₆, —NR₅C(═O)R₆, —NR₅SO₂R₅, —SO₂R₅, —S(═O)₂NR₅R₆, —POR₅R₆,—C₃₋₆carbocyclic, 3- 6 membered heterocyclic, —C₆₋₁₀aryl, or 5-10membered heteroaryl, each hydrogen in R₁₁, R₁₂, R₁₃, R₁₄ or R₁₅ isindependently optionally substituted with 1, 2, 3, 4, 5 or 6substituent(s) selected from halogen, —C₁₋₆alkyl, —C₂₋₆alkenyl,—C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN, oxo, —OC₁₋₆alkyl, —SC₁₋₆alkyl,—NR₅R₆, —C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅, —C(═O)NR₅R₆, —NR₅C(═O)R₆,—NR₅SO₂R₆, —SO₂R₅, —S(═O)₂NR₅R₆, —POR₅R₆, —C₃₋₆carbocyclic, 3-6 memberedheterocyclic, —C₆₋₁₀aryl, or 5-10 membered heteroaryl; each heterocyclicor heteroaryl at each occurrence independently contains 1, 2, 3 or 4heteroatom(s) selected from N, O, S, S═O or S(═O)₂;

R₂₁ or R₂₂ is independently selected from hydrogen, halogen, —C₁₋₆alkyl,—C₂₋₆alkenyl, —C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN, —OC₁₋₆alkyl,—SC₁₋₆alkyl, —NR₅R₆, —C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅, —C(═O)NR₅R₆,—NR₅C(═O)R₆, —NR₅SO₂R₅, —SO₂R₅, —S(═O)₂NR₅R₆, —POR₅R₆, —C₃₋₆carbocyclic,3- 6 membered heterocyclic, —C₆₋₁₀aryl, or 5-10 membered heteroaryl,each hydrogen in R₂₁ or R₂₂ is independently optionally substituted with1, 2, 3, 4, 5 or 6 substituent(s) selected from halogen, —C₁₋₆alkyl,—C₂₋₆alkenyl, —C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN, oxo, —OC₁₋₆alkyl,—SC₁₋₆alkyl, —NR₅R₅, —C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅, —C(═O)NR₅R₆,—NR₅C(═O)R₆, —S(═O)₂NR₅R₆, —POR₅R₆, —C₃₋₆carbocyclic, 3-6 memberedheterocyclic, —C₆₋₁₀aryl, or 5-10 membered heteroaryl; each heterocyclicor heteroaryl at each occurrence independently contains 1, 2, 3 or 4heteroatom(s) selected from N, O, S, S═O or S(═O)₂; R₃ is selected from—C₁₋₁₄alkyl, —C₂₋₁₄alkenyl, —C₂₋₁₄alkynyl, —C₆₋₁₀aryl, 5-10 memberedheteroaryl, 3-14 membered heterocyclic, —C₃₋₁₄carbocyclic,

each ring C at each occurrence is independently selected from a C₃₋₁₄carbocyclic or 3-14 membered heterocyclic ring, each ring D at eachoccurrence is independently selected from a C₆₋₁₀ aryl or 5-10 memberedheteroaryl ring, each hydrogen in R₃ is independently optionallysubstituted with 1, 2, 3, 4, 5 or 6 R₃₁; each heterocyclic or heteroarylat each occurrence independently contains 1, 2, 3 or 4 heteroatomsselected from N, O, S, S═O or S(═O)₂;

Each R₃₁ at each occurrence is independently selected from halogen,—C₁₋₆alkyl, —C₂₋₆alkenyl, —C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN,—OC₁₋₆alkyl, —SC₁₋₆alkyl, —NR₅R₆, —C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅,—C(═O)NR₅R₆, —NR₅C(═O)R₆, —NR₅SO₂R₆, —SO₂R₅, —S(═O)₂NR₅R₆, —POR₅R₆,—C₃₋₆carbocyclic, 3- 6 membered heterocyclic, —C₆₋₁₀aryl, or 5-10membered heteroaryl, each hydrogen in R₃₁ is independently optionallysubstituted with 1, 2, 3, 4, 5 or 6 substituent(s) selected fromhalogen, —C₁₋₆alkyl, —C₂₋₆alkenyl, —C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN,oxo, —OC₁₋₆alkyl, —SC₁₋₆alkyl, —NR₅R₆, —C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅,—C(═O)NR₅R₆, —NR₅C(═O)R₆, —S(═O)₂NR₅R₆, —POR₅R₆, —C₃₋₆carbocyclic, 3-6membered heterocyclic, —C₆₋₁₀aryl, or 5-10 membered heteroaryl; eachheterocyclic or heteroaryl at each occurrence independently contains 1,2, 3 or 4 heteroatom(s) selected from N, O, S, S═O or S(═O)₂;

R₄ is selected from

Each G₁, G₂, G₃ or G₄ at each occurrence is independently selected fromN or CH;

Each n1, n2, n3, n4 or n5 at each occurrence is independently selectedfrom 0, 1, 2, 3, 4, 5 or 6, provided that n1 and n2 is not 0 at the sametime, n3 and n4 is not 0 at the same time;

Each hydrogen in

is independently optionally substituted with 1R₄₂, 2R₄₂, 3R₄₂, 4R₄₂,5R₄₂ or 6R₄₂;

Each R₄₁ at each occurrence is independently selected from

Each of Q at each occurrence is independently selected from C(═O),NR₅C(═O), S(═O)₂ or NR₅S(═O)₂;

in

is selected from

or

;

when

is

, R_(4a), R_(4b) or R_(4c) is independently selected from hydrogen,halogen, —C₁₋₆alkyl, —C₂₋₆alkenyl, —C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN,—OC₁₋₆alkyl, —SC₁₋₆alkyl, —NR₅R₅, —C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅,—C(═O)NR₅R₆, —NR₅C(═O)R₆, —NR₅SO₂R₆, —SO₂R₅, —S(═O)₂NR₅R₆, —POR₅R₆,—C₃₋₆carbocyclic, 3-6 membered heterocyclic, —C₆₋₁₀aryl, or 5-10membered heteroaryl, each hydrogen in R_(4a), R_(4b) or R_(4c) isindependently optionally substituted with 1, 2, 3, 4, 5 or 6substituents selected from halogen, —C₁₋₆alkyl, —C₂₋₆alkenyl,—C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN, oxo, —OC₁₋₆alkyl, —SC₁₋₆alkyl,—NR₅R₅, —C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅, —C(═O)NR₅R₆, —NR₅C(═O)R₆,—NR₅SO₂R₆, —SO₂R₅, —S(═O)₂NR₅R₆, —POR₅R₆, —C₃₋₆carbocyclic, 3-6 memberedheterocyclic, —C₆₋₁₀aryl, or 5-10 membered heteroaryl; each heterocyclicor heteroaryl at each occurrence independently contains 1, 2, 3 or 4heteroatom(s) selected from N, O, S, S═O or S(═O)₂; or

when

is

, R_(4a) is selected from hydrogen, halogen, —C₁₋₆alkyl, —C₂₋₆alkenyl,—C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN, —OC₁₋₆alkyl, —SC₁₋₆alkyl, —NR₅R₆,—C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅, —C(═O)NR₅R₆, —NR₅C(═O)R₆, —NR₅SO₂R₅,—SO₂R₅, —S(═O)₂NR₅R₆, —POR₅R₆, —C₃₋₆carbocyclic, 3- 6 memberedheterocyclic, —C₆₋₁₀aryl, or 5-10 membered heteroaryl, each hydrogen inR_(4a) is independently optionally substituted with 1, 2, 3, 4, 5 or 6substituents selected from halogen, —C₁₋₆alkyl, —C₂₋₆alkenyl,—C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN, oxo, —OC₁₋₆alkyl, —SC₁₋₆alkyl,—NR₅R₆, —C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅, —C(═O)NR₅R₆, —NR₅C(═O)R₆,—NR₅SO₂R₆, —SO₂R₅, —S(═O)₂NR₅R₆, —POR₅R₆, —C₃₋₆carbocyclic, 3-6 memberedheterocyclic, —C₆₋₁₀aryl, or 5-10 membered heteroaryl; and R_(4b) andR_(4c) together with the carbon which they both attach to form a C₃₋₁₀carbocyclic ring or a 3-10 membered heterocyclic ring, each hydrogen inthe C₃₋₁₀ carbocyclic ring or the 3-10 membered heterocyclic ring isoptionally substituted by 1, 2, 3, 4, 5 or 6 substituents selected fromhalogen, —C₁₋₆alkyl, —C₂₋₆alkenyl, —C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN,oxo, —OC₁₋₆alkyl, —SC₁₋₆alkyl, —NR₅R₆, —C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅,—C(═O)NR₅R₆, —NR₅C(═O)R₆, —NR₅SO₂R₆, —SO₂R₅, —S(═O)₂NR₅R₆, —POR₅R₆,—C₃₋₆carbocyclic, 3-6 membered heterocyclic, —C₆₋₁₀aryl, or 5-10membered heteroaryl; each heterocyclic or heteroaryl at each occurrenceindependently contains 1, 2, 3 or 4 heteroatom(s) selected from N, O, S,S═O or S(═O)₂; or

when

is

, R_(4a) and R_(4c) with the carbon they respectively attach to form aC₃₋₁₀ carbocyclic ring or a 3-10 membered heterocyclic ring, eachhydrogen in the C₃₋₁₀ carbocyclic ring or the 3-10 membered heterocyclicring is optionally substituted by 1, 2, 3, 4, 5 or 6 substituentsselected from halogen, —C₁₋₆alkyl, —C₂₋₆alkenyl, —C₂₋₆alkynyl,heteroC₂₋₆alkyl, —CN, oxo, —OC₁₋₆alkyl, —SC₁₋₆alkyl, —NR₅R₅, —C(═O)R₅,—C(═O)OR₅, —OC(═O)R₅, —C(═O)NR₅R₆, —NR₅C(═O)R₆, —NR₅SO₂R₆, —SO₂R₅,—S(═O)₂NR₅R₆, —POR₅R₆, —C₃₋₆carbocyclic, 3-6 membered heterocyclic,—C₆₋₁₀aryl, or 5-10 membered heteroaryl; and R_(4b) is selected fromhydrogen, halogen, —C₁₋₆alkyl, —C₂₋₆alkenyl, —C₂₋₆alkynyl,heteroC₂₋₆alkyl, —CN, —OC₁₋₆alkyl, —SC₁₋₆alkyl, —NR₅R₆, —C(═O)R₅,—C(═O)OR₅, —OC(═O)R₅, —C(═O)NR₅R₆, —NR₅C(═O)R₆, —NR₅SO₂R₆, —SO₂R₅,—S(═O)₂NR₅R₆, —POR₅R₆, —C₃₋₆carbocyclic, 3-6 membered heterocyclic,—C₆₋₁₀aryl, or 5-10 membered heteroaryl, each hydrogen in R_(4b) isindependently optionally substituted with 1, 2, 3, 4, 5 or 6substituents selected from halogen, —C₁₋₆alkyl, —C₂₋₆alkenyl,—C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN, oxo, —OC₁₋₆alkyl, —SC₁₋₆alkyl,—NR₅R₆, —C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅, —C(═O)NR₅R₆, —NR₅C(═O)R₆,—NR₅SO₂R₆, —SO₂R₅, —S(═O)₂NR₅R₆, —POR₅R₆, —C₃₋₆carbocyclic, 3-6 memberedheterocyclic, —C₆₋₁₀aryl, or 5-10 membered heteroaryl; each heterocyclicor heteroaryl at each occurrence independently contains 1, 2, 3 or 4heteroatom(s) selected from N, O, S, S═O or S(═O)₂;

when

is

, R_(4a) is absent, R_(4b) is absent, R_(4c) is selected from hydrogen,halogen, —C₁₋₆alkyl, —C₂₋₆alkenyl, —C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN,—OC₁₋₆alkyl, —SC₁₋₆alkyl, —NR₅R₆, —C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅,—C(═O)NR₅R₆, —NR₅C(═O)R₆, —NR₅SO₂R₆, —SO₂R₅, —S(═O)₂NR₅R₆, —POR₅R₆,—C₃₋₆carbocyclic, 3-6 membered heterocyclic, —C₆₋₁₀aryl, or 5-10membered heteroaryl, each hydrogen in R_(4c) is independently optionallysubstituted with 1, 2, 3, 4, 5 or 6 substituents selected from halogen,—C₁₋₆alkyl, —C₂₋₆alkenyl, —C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN, oxo,—OC₁₋₆alkyl, —SC₁₋₆alkyl, —NR₅R₆, —C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅,—C(═O)NR₅R₆, —NR₅C(═O)R₆, —NR₅SO₂R₆, —SO₂R₅, —S(═O)₂NR₅R₆, —POR₅R₆,—C₃₋₆carbocyclic, 3-6 membered heterocyclic, —C₆₋₁₀aryl, or 5-10membered heteroaryl; each heterocyclic or heteroaryl at each occurrenceindependently contains 1, 2, 3 or 4 heteroatom(s) selected from N, O, S,S═O or S(═O)₂;

R_(4a) is halogen;

Each R₄₂ at each occurrence is independently selected from halogen,—C₁₋₆alkyl, —C₂₋₆alkenyl, —C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN, oxo,—OC₁₋₆alkyl, —SC₁₋₆alkyl, —NR₅R₆, —C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅,—C(═O)NR₅R₆, —NR₅C(═O)R₆, —NR₅SO₂R₆, —SO₂R₅, —S(═O)₂NR₅R₆, —POR₅R₆,—C₃₋₆carbocyclic, 3-6 membered heterocyclic, —C₆₋₁₀aryl, or 5-10membered heteroaryl, each hydrogen in R₄₂ is independently optionallysubstituted with 1, 2, 3, 4, 5 or 6 substituents selected from halogen,—C₁₋₆alkyl, —C₂₋₆alkenyl, —C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN, oxo,—OC₁₋₆alkyl, —SC₁₋₆alkyl, —NR₅R₆, —C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅,—C(═O)NR₅R₆, —NR₅C(═O)R₆, —NR₅SO₂R₆, —SO₂R₅, —S(═O)₂NR₅R₆, —POR₅R₆,—C₃₋₆carbocyclic, 3-6 membered heterocyclic, —C₆₋₁₀aryl, or 5-10membered heteroaryl; or

Two R₄₂ together with the atom which they both or respectively attach toform a C₃₋₆ carbocyclic or 3-6 membered heterocyclic ring, each hydrogenin the C₃₋₆ carbocyclic or 3-6 membered heterocyclic ring isindependently optionally substituted with 1, 2, 3, 4, 5 or 6substituents selected from halogen, —C₁₋₆alkyl, —C₂₋₆alkenyl,—C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN, oxo, —OC₁₋₆alkyl, —SC₁₋₆alkyl,—NR₅R₆, —C(═O)R₅, —C(═O)OR₅, —OC(═O)R₅, —C(═O)NR₅R₆, —NR₅C(═O)R₅,—NR₅SO₂R₆, —SO₂R₅, —S(═O)₂NR₅R₆, —PO(R₅)₂, —C₃₋₆carbocyclic, 3-6membered heterocyclic, —C₆₋₁₀aryl, or 5-10 membered heteroaryl; eachheterocyclic and heteroaryl at each occurrence independently contains 1,2, 3 or 4 heteroatom(s) selected from N, O, S, S═O or S(═O)₂;

Each R₅ and R₆ at each occurrence is independently selected fromhydrogen or —C₁₋₆alkyl; or

R₅ and R₆ together with the atom which they both or respectively attachto form a 3-10 membered heterocyclic ring, the 3-10 memberedheterocyclic ring is optionally further contains 1, 2, 3 or 4heteroatoms selected from N, O, S, S(═O) or S(═O)₂, and each hydrogen inthe 3-10 membered heterocyclic ring is independently optionallysubstituted with 1R₅₁, 2R₅₁, 3R₅₁, 4R₅₁, 5R₅₁ or 6R₅₁;

Each R₅₁ is selected from halogen, —C₁₋₆alkyl, —C₂₋₆alkenyl,—C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN, —OC₁₋₆alkyl, —SC₁₋₆alkyl, —NR₇R₈,—C(═O)R₇, —C(═O)OR₇, —OC(═O)R₇, —C(═O)NR₇R₅, —NR₇C(═O)R₅, —NR₇SO₂R₈,—SO₂R₇, —S(═O)₂NR₇R₅, —POR₇R₈, —C₃₋₆carbocyclic, 3-6 memberedheterocyclic, —C₆₋₁₀aryl, or 5-10 membered heteroaryl, each hydrogen inR₅₁ is independently optionally substituted with 1, 2, 3, 4, 5 or 6substituent(s) selected from halogen, —C₁₋₆alkyl, —C₂₋₆alkenyl,—C₂₋₆alkynyl, heteroC₂₋₆alkyl, —CN, oxo, —OC₁₋₆alkyl, —SC₁₋₆alkyl,—NR₇R₈, —C(═O)R₇, —C(═O)OR₇, —OC(═O)R₇, —C(═O)NR₇R₅, —NR₇C(═O)R₅,—NR₇SO₂R₈, —SO₂R₇, —S(═O)₂NR₇R₅, —POR₇R₈, —C₃₋₆carbocyclic, 3-6 memberedheterocyclic, —C₆₋₁₀aryl, or 5-10 membered heteroaryl; each heterocyclicor heteroaryl at each occurrence independently contains 1, 2, 3 or 4heteroatom(s) selected from N, O, S, S═O or S(═O)₂;

Each R₇ and R₈ at each occurrence is independently selected fromhydrogen or —C₁₋₆alkyl.

In some embodiments, there is provided a compound of formula (I), astereoisomer thereof, an atropisomer thereof, a pharmaceuticallyacceptable salt thereof, a pharmaceutically acceptable salt of thestereoisomer thereof or a pharmaceutically acceptable salt of theatropisomer thereof:

Wherein:

R₁₁, R₁₂, R₁₃, R₁₄ or R₁₅ is independently selected from —OH; halogen;—NR_(a)R_(b); —C₁₋₆alkyl; —OC₁₋₆alkyl; —C₁₋₆alkylene-OH;—C₁₋₆alkylene-O—C₁₋₆alkyl; —C₁₋₆alkyl substituted with halogen, —NH₂,—CN or —OH; —O—C₁₋₆alkyl substituted with halogen, —NH₂, —CN or —OH;—O—C₁₋₆alkyl; —SO₂R_(a); —CN; —C(═O)NR_(a)R_(b); —C(═O)R_(a);—OC(═O)R_(a); —C(═O)OR_(a); or —C₃₋₆carbocyclic;

R_(a) or R_(b) is independently selected from hydrogen or —C₁₋₆alkyl;

R₂₁ is selected from hydrogen; halogen; —C₁₋₆alkyl; —C₁₋₆alkylsubstituted with halogen, —NH₂, —CN or —OH; —C₂₋₆alkenyl; or—C₃₋₆carbocyclic;

R₂₂ is selected from hydrogen; halogen; —C₁₋₆alkyl; —C₁₋₆alkylsubstituted with halogen, —NH₂, —CN or —OH; —C₂₋₆alkenyl; or—C₃₋₆carbocyclic;

R₃ is selected from —C₆₋₁₀aryl or 5-10 membered heteroaryl, each of 5-10membered heteroaryl at each occurrence independently contains 1, 2, 3 or4 heteroatoms selected from N, O or S, each hydrogen in the —C₆₋₁₀arylor 5-10 membered heteroaryl at each occurrence is independentlyoptionally substituted with 1R₃₁, 2R₃₁, 3R₃₁, 4R₃₁, 5R₃₁ or 6 R₃₁;

Each R₃₁ at each occurrence is independently selected from halogen,—C₁₋₆alkyl, —CN, —OH, —O—C₁₋₆alkyl, —NH₂, —NH(C₁₋₆alkyl), —N(C₁₋₆alkyl)₂or —C₃₋₆carbocyclic;

R₄ is

each hydrogen in the

is independently optionally substituted with 1R₄₂, 2R₄₂, 3R₄₂, 4R₄₂,5R₄₂ or 6 R₄₂;

n1 or n2 is independently selected from 1, 2, 3, 4, 5 or 6;

R₄₁ is

R_(4a), R_(4b) or R_(4c) is independently selected from hydrogen,halogen, —C₁₋₆alkyl or —C₁₋₆alkylene-N(C₁₋₆alkyl)₂;

Each R₄₂ at each occurrence is independently selected from —C₁₋₆alkyl;—C₁₋₆alkylene-CN or —C₁₋₆alkyl substituted with halogen.

In some embodiments, R₁₁, R₁₂, R₁₃, R₁₄ or R₁₅ is independently selectedfrom —OH; —F; —Cl; —Br; —NR_(a)R_(b); —C₁₋₃alkyl; —OC₁₋₃alkyl;—C₁₋₃alkylene-OH; —C₁₋₃alkylene-O—C₁₋₃alkyl; —C₁₋₃alkyl substituted with—F or —Cl; —O—C₁₋₃alkyl substituted with —F or —Cl; —SO₂R_(a); —CN;—C(═O)NR_(a)R_(b); —C(═O)R_(a); —OC(═O)R_(a); —C(═O)OR_(a); 3-memberedcarbocyclic; 4-membered carbocyclic; 5-membered carbocyclic or6-membered carbocyclic;

R_(a) or R_(b) is independently selected from hydrogen or —C₁₋₃alkyl.

In some embodiments, R₁₁, R₁₂, R₁₃, R₁₄ or R₁₅ is independently selectedfrom —OH, —F, —Cl, —NH₂, —NHCH₃, —N(CH₃)₂, —CH3, —CH₂CH₃, —CH₂CH₂CH₃,—CH(CH₃)₂, —OCH₃, —OCH₂CH₃, —OCH₂CH₂CH₃, —OCH(CH₃)₂, —CH₂OH, —CH₂CH₂OH,—CH₂OCH₃, —CHF₂, —CH₂F, —CF₃, —OCH₂F, —OCHF₂, —OCF₃, —SO₂CH₃, —CN,—C(═O)NH₂, —C(═O)CH₃, —OC(═O)CH₃, —C(═O)OCH₃ or 3-membered carbocyclic.

In some embodiments, R₁₁, R₁₂, R₁₃, R₁₄ or R₁₅ is independently selectedfrom —OH, —F, —Cl, —NH₂, —CH₃ or —CF₃.

In some embodiments, the

in the formula (I) is selected from:

In some embodiments, R₂₁ is selected from hydrogen; —F; —Cl; —Br;—C₁₋₃alkyl; —C₁₋₃alkyl substituted with —F or —Cl; —C₂₋₃alkenyl; or—C₃₋₆carbocyclic.

In some embodiments, R₂₁ is selected from hydrogen; —F; —Cl; methyl;ethyl; propyl; isopropyl; methyl substituted with —F; ethyl substitutedwith —F; propyl substituted with —F; isopropyl substituted with —F;ethenyl; propenyl; 3 membered carbocyclic; 4 membered carbocyclic; 5membered carbocyclic; or 6 membered carbocyclic.

In some embodiments, R₂₁ is —C₁.

In some embodiments, R₂₂ is selected from hydrogen; —F; —Cl; —Br;—C₁₋₃alkyl; —C₁₋₃alkyl substituted with —F or —Cl; —C₂₋₃alkenyl; or—C₃₋₆carbocyclic.

In some embodiments, R₂₂ is selected from hydrogen; —F; —Cl; methyl;ethyl; propyl; isopropyl; methyl substituted with —F; ethyl substitutedwith —F; propyl substituted with —F; isopropyl substituted with —F;ethenyl; propenyl; 3 membered carbocyclic; 4 membered carbocyclic; 5membered carbocyclic; or 6 membered carbocyclic.

In some embodiments, R₂₂ is hydrogen.

In some embodiments, R₃ is selected from phenyl, naphthyl, 5 memberedheteroaryl, 6 membered heteroaryl, 7 membered heteroaryl, 8 memberedheteroaryl, 9 membered heteroaryl or 10 membered heteroaryl, eachheteroaryl at each occurrence independently contains 1, 2 or 3heteroatoms selected from N or O, each hydrogen in R₃ at each occurrenceis independently optionally substituted with 1R₃₁, 2R₃₁, 3R₃₁, 4R₃₁ or5R₃₁.

In some embodiments, R₃ is selected from phenyl or 6 memberedheteroaryl, the heteroaryl contains 1 or 2 heteroatoms selected from N,each hydrogen in the phenyl or 6 membered heteroaryl at each occurrenceis independently optionally substituted by 1R₃₁, 2R₃₁, 3R₃₁ or 4R₃₁.

In some embodiments, R₃ is selected from

each hydrogen in the R₃ at each occurrence is independently optionallysubstituted by 1R₃₁, 2R₃₁, 3R₃₁ or 4R₃₁.

In some embodiments, each R₃₁ at each occurrence is independentlyselected from —F, —Cl, —Br, —C₁₋₃alkyl, —CN, —OH, —O—C₁₋₃alkyl, —NH₂,—NH(C₁₋₃alkyl), —N(C₁₋₃alkyl)₂ or —C₃₋₆carbocyclic.

In some embodiments, each R₃₁ at each occurrence is independentlyselected from —F, —Cl, methyl, ethyl, propyl, isopropyl, —CN, —OH,methoxy, ethoxy, propoxy, isopropoxy, —NH₂, —NHCH₃, —NHCH₂CH₃,—NH(CH₂CH₂CH₃), —NH(CH(CH₃)₂), —N(CH₃)₂, —N(CH₂CH₃)₂, —N(CH₃)(CH₂CH₃), 3membered carbocyclic, 4 membered carbocyclic, 5 membered carbocyclic or6 membered carbocyclic.

In some embodiments, each R₃₁ at each occurrence is independentlyselected from methyl or isopropyl.

In some embodiments, R₃ is selected from

In some embodiments, R₃ is

In some embodiments, R₄ is

each hydrogen in the

is independently optionally substituted with 1R₄₂, 2R₄₂, 3R₄₂ or 4R₄₂;

n1 is selected from 1, 2 or 3;

n2 is selected from 1, 2 or 3.

In some embodiments, R₄ is selected from

each hydrogen in the

is independently optionally substituted with 1R₄₂ or 2R₄₂;

n1 is selected from 1 or 2;

n2 is selected from 1 or 2.

In some embodiments, R₄ is

each hydrogen in the

is independently optionally substituted with 1R₄₂ or 2R₄₂.

In some embodiments, R₄₁ is

R_(4a), R_(4b), or R_(4c) is independently selected from hydrogen, —F,—Cl, —Br, —C₁₋₃alkyl or —C₁₋₃alkylene-N(C₁₋₃alkyl)₂.

In some embodiments, R_(4a), R_(4b) or R_(4c) is independently selectedfrom hydrogen, —F, —Cl, methyl, ethyl, propyl, isopropyl, —CH₂—N(CH₃)₂,—CH₂—N(CH₂CH₃)₂ or —CH₂—N(CH₃)(CH₂CH₃).

In some embodiments, R_(4a), R_(4b) or R_(4c) is independently selectedfrom hydrogen, —F, methyl or —CH₂—N(CH₃)₂.

In some embodiments, R_(4a) is selected from hydrogen or —F; R_(4b) ishydrogen; R_(4c) is selected from hydrogen or —CH₂—N(CH₃)₂.

In some embodiments, R₄₁ is selected from:

In some embodiments, R₄ is selected from

each hydrogen in the

at each occurrence is independently optionally substituted with 1R₄₂ or2R₄₂.

In some embodiments, each R₄₂ at each occurrence is independentlyselected from —C₁₋₃alkyl; —C₁₋₃alkylene-CN; or —C₁₋₃alkyl substitutedwith —F or —C₁.

In some embodiments, each R₄₂ at each occurrence is independentlyselected from methyl; ethyl; propyl; isopropyl; -methylene-CN;-ethylene-CN; -propylene-CN; methyl substituted with —F; ethylsubstituted with —F; propyl substituted with —F; or isopropylsubstituted with —F.

In some embodiments, each R₄₂ at each occurrence is independentlyselected from methyl; ethyl; -methylene-CN or methyl substituted with—F.

In some embodiments, each R₄₂ at each occurrence is independentlyselected from —CH₃, —CH₂CH₃, —CH₂CN, —CHF₂ or —CF₃.

In some embodiments, R₄ is selected from

In some embodiments, R₄ is selected from:

In some embodiments, the compound is selected from:

In another aspect, provided herein is a method for preparing thecompound of formula (I), a stereoisomer thereof, an atropisomer thereof,a pharmaceutically acceptable salt thereof, a pharmaceuticallyacceptable salt of the stereoisomer thereof or a pharmaceuticallyacceptable salt of the atropisomer thereof, the method comprises acoupling reaction between a compound of formula (II) and a compound offormula (III) according to the following reaction Scheme 1 or between acompound of formula (II′) and a compound of formula (III′) according tothe following reaction Scheme 2 catalyzed by a transition metalpalladium or nickel reagent:

Wherein

the L in the compound of formula (III) or formula (II′) is a leavinggroup; preferably, the leaving group is selected from halogen,—OS(O)₂CF₃ or —OTs; more preferably, the halogen is selected from —F,—Cl, —Br, or —I; more preferably, the leaving group is —Cl or —Br;

the X in the compound of formula (II) or formula (III′) is selected fromboronic acid, borate ester or organotin; more preferably, the X isselected from

preferably, the coupling reaction is Suzuki coupling reaction or Stillecoupling reaction;

preferably, the coupling reaction is catalyzed by the transition metalpalladium reagent; more preferably, the transition metal palladiumreagent is Pd(PPh₃)₄.

In another aspect, provided herein is a pharmaceutical compositioncomprising the compound of formula (I), a stereoisomer thereof, anatropisomer thereof, a pharmaceutically acceptable salt thereof, apharmaceutically acceptable salt of the stereoisomer thereof or apharmaceutically acceptable salt of the atropisomer thereof of thepresent invention, and at least one pharmaceutically acceptableexcipient. In some embodiments, the said compound in a weight ratio tothe said excipient within the range from about 0.0001 to about 10.

In another aspect, provided herein is use of the compound of formula(I), a stereoisomer thereof, an atropisomer thereof, a pharmaceuticallyacceptable salt thereof, a pharmaceutically acceptable salt of thestereoisomer thereof or a pharmaceutically acceptable salt of theatropisomer thereof of the present invention; or the pharmaceuticalcomposition of the present invention for the manufacture of a medicamentfor the treatment of diseases or conditions related to KRAS mutantprotein. In some embodiments, the diseases or conditions related to KRASmutant protein is the diseases or conditions related to KRAS G12C mutantprotein. In some embodiments, the diseases or conditions related to KRASG12C mutant protein is cancer related to KRAS G12C mutant protein. Insome embodiments, the cancer is selected from blood cancer, pancreaticcancer, colon cancer, rectal cancer, colorectal cancer or lung cancer.In some embodiments, the blood cancer is selected from acute myeloidleukemia or acute lymphocytic leukemia; the lung cancer is selected fromnon-small cell lung cancer or small cell lung cancer.

In another aspect, provided herein is a method of treating a subjecthaving a diseases or conditions related to KRAS mutant protein, saidmethod comprising administering to the subject a therapeuticallyeffective amount of the compound of formula (I), a stereoisomer thereof,an atropisomer thereof, a pharmaceutically acceptable salt thereof, apharmaceutically acceptable salt of the stereoisomer thereof or apharmaceutically acceptable salt of the atropisomer thereof of thepresent invention; or the pharmaceutical composition of the presentinvention. In some embodiments, the diseases or conditions related toKRAS mutant protein is the diseases or conditions related to KRAS G12Cmutant protein. In some embodiments, the diseases or conditions relatedto KRAS G12C mutant protein is cancer related to KRAS G12C mutantprotein. In some embodiments, the cancer is selected from blood cancer,pancreatic cancer, colon cancer, rectal cancer, colorectal cancer orlung cancer. In some embodiments, the blood cancer is selected fromacute myeloid leukemia or acute lymphocytic leukemia; the lung cancer isselected from non-small cell lung cancer or small cell lung cancer.

Definition

The term “halogen” or “halo”, as used herein, unless otherwiseindicated, means fluoro, chloro, bromo or iodo. The preferred halogengroups include —F, —Cl and —Br. The preferred halogen groups include —Fand —Cl.

The term “alkyl”, as used herein, unless otherwise indicated, includessaturated monovalent hydrocarbon radicals having straight or branched.For example, alkyl radicals include methyl, ethyl, propyl, isopropyl,n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, 3-(2-methyl)butyl,2-pentyl, 2-methylbutyl, neopentyl, n-hexyl, 2-hexyl and 2-methylpentyl.Similarly, C₁-6, as in C₁₋₆alkyl is defined to identify the group ashaving 1, 2, 3, 4, 5 or 6 carbon atoms in a linear or branchedarrangement.

The term “alkylene” means a difunctional group obtained by removal of ahydrogen atom from an alkyl group that is defined above. For example,methylene (i.e., —CH₂—), ethylene (i.e., —CH₂—CH₂— or —CH(CH₃)—) andpropylene (i.e., —CH₂—CH₂— CH₂—, —CH(—CH₂—CH₃)— or —CH₂—CH(CH₃)—).

The term “alkenyl” means a straight or branch-chained hydrocarbonradical containing one or more double bonds and typically from 2 to 20carbon atoms in length. For example, “C₂₋₆alkenyl” contains from 2 to 6carbon atoms. Alkenyl group include, but are not limited to, forexample, ethenyl, propenyl, butenyl, 2-methyl-2-buten-1-yl, heptenyl,octenyl and the like.

The term “alkynyl” contains a straight or branch-chained hydrocarbonradical containing one or more triple bonds and typically from 2 to 20carbon atoms in length. For example, “C₂₋₆alkynyl” contains from 2 to 6carbon atoms. Representative alkynyl groups include, but are not limitedto, for example, ethynyl, 1-propynyl, 1-butynyl, heptynyl, octynyl andthe like.

The term “alkoxy” radicals are oxygen ethers formed from the previouslydescribed alkyl groups.

The term “aryl” or “aryl ring”, as used herein, unless otherwiseindicated, refers to an unsubstituted or substituted mono or polycyclicaromatic ring system only containing carbon ring atoms. The preferredaryls are mono cyclic or bicyclic 6-10 membered aromatic ring systems.Phenyl and naphthyl are preferred aryls.

The term “heterocyclic” or “heterocyclic ring”, as used herein, unlessotherwise indicated, refers to unsubstituted and substituted mono orpolycyclic non-aromatic ring system containing one or more ringheteroatom(s), which comprising moncyclic heterocyclic (ring), bicyclicheterocyclic (ring), bridged heterocyclic (ring), fused heterocyclic(ring) or sipro heterocyclic (ring). Preferred heteroatoms include N, O,and S, including N-oxides, sulfur oxides, and dioxides. Preferably theheterocyclic (ring) is three to ten membered and is either fullysaturated or has one or more degrees of unsaturation. Multiple degreesof substitution, preferably one, two or three, are included within thepresent definition of heterocyclic (ring). Examples of such heterocyclicgroups include, but are not limited to azetidinyl, pyrrolidinyl,piperidinyl, piperazinyl, oxopiperazinyl, oxopiperidinyl, oxoazepinyl,azepinyl, tetrahydrofuranyl, dioxolanyl, tetrahydroimidazolyl,tetrahydrothiazolyl, tetrahydrooxazolyl, tetrahydropyranyl, morpholinyl,thiomorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone andoxadiazolyl.

The term “heteroaryl” or “heteroaryl ring”, as used herein, unlessotherwise indicated, represents an aromatic ring system containingcarbon(s) and at least one heteroatom. Heteroaryl or heteroaryl ring maybe monocyclic or polycyclic, substituted or unsubstituted. A monocyclicheteroaryl group may have 1 to 4 heteroatoms in the ring, while apolycyclic heteroaryl may contain 1 to 10 hetero atoms. A polycyclicheteroaryl ring may contain fused, spiro or bridged ring junction, forexample, bycyclic heteroaryl is a polycyclic heteroaryl. Bicyclicheteroaryl rings may contain from 8 to 12 member atoms. Monocyclicheteroaryl rings may contain from 5 to 8 member atoms (cabons andheteroatoms). Examples of heteroaryl groups include, but are not limitedto thienyl, furanyl, imidazolyl, isoxazolyl, oxazolyl, pyrazolyl,pyrrolyl, thiazolyl, thiadiazolyl, triazolyl, pyridyl, pyridazinyl,indolyl, azaindolyl, indazolyl, benzimidazolyl, benzofuranyl,benzothienyl, benzisoxazolyl, benzoxazolyl, benzopyrazolyl,benzothiazolyl, benzothiadiazolyl, benzotriazolyl adeninyl, quinolinylor isoquinolinyl.

The term “carbocyclic” or “carbocyclic ring” refers to a substituted orunsubstituted monocyclic ring, bicyclic ring, bridged ring, fused ring,sipiro ring non-aromatic ring system only containing carbon atoms. Thecarbocyclic (ring) contain cycloalkyl without substituted degrees andcarbocyclic with one or more substituted degrees. Exemplary “cycloalkyl”groups includes but not limited to, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl and so on.

The term “oxo” refers to oxygen atom together with the attached carbonatom forms the group

The term “—C₁₋₆alkylene-N(C₁₋₆alkyl)₂” refers to the —C₁₋₆alkyl asdefined above substituted by —N(C₁₋₆alkyl)₂.

The term “—C₁₋₆alkylene-CN” refers to the —C₁₋₆alkyl as defined abovesubstituted by —CN.

The term “heteroalkyl” refers to the presence of heteroatoms between anytwo carbon atoms in the alkyl group as defined above, such as N or Oatoms. For example, “heteroC₂₋₆alkyl” means that there are N atom or Oatom between any two carbon atoms in the C₂₋₆ alkyl group, including butnot limited to —CH₂OCH₃, —CH₂CH₂OCH₃, —CH₂NHCH₃, or —CH₂N(CH₃)₂ and thelike.

The term “composition”, as used herein, is intended to encompass aproduct comprising the specified ingredients in the specified amounts,as well as any product which results, directly or indirectly, fromcombinations of the specified ingredients in the specified amounts.Accordingly, pharmaceutical compositions containing the compounds of thepresent invention as the active ingredient as well as methods ofpreparing the instant compounds are also part of the present invention.

The compounds of the present invention may also be present in the formof pharmaceutically acceptable salt(s). For use in medicine, the saltsof the compounds of this invention refer to non-toxic “pharmaceuticallyacceptable salt(s)”. The pharmaceutically acceptable salt forms includepharmaceutically acceptable acidic/anionic or basic/cationic salts. Thepharmaceutically acceptable acidic/anionic salt generally takes a formin which the basic nitrogen is protonated with an inorganic or organicacid.

The present invention includes within its scope the prodrugs of thecompounds of this invention. In general, such prodrugs will befunctional derivatives of the compounds that are readily converted invivo into the required compound. Thus, in the methods of treatment ofthe present invention, the term “administering” shall encompass thetreatment of the various disorders described with the compoundspecifically disclosed or with a compound which may not be specificallydisclosed, but which converts to the specified compound in vivo afteradministration to the subject. Conventional procedures for the selectionand preparation of suitable prodrug derivatives are described, forexample, in “Design of Prodrugs”, ed. H. Bundgaard, Elsevier, 1985.

It is intended that the definition of any substituent or variable at aparticular location in a molecule be independent of its definitionselsewhere in that molecule. It is understood that substituents andsubstitution patterns on the compounds of this invention can be selectedby one of ordinary skill in the art to provide compounds that arechemically stable and that can be readily synthesized by techniques knowin the art as well as those methods set forth herein.

The present invention includes compounds described can contain one ormore asymmetric centers and may thus give rise to diastereomers andoptical isomers. The present invention includes all such possiblediastereomers as well as their racemic mixtures, their substantiallypure resolved enantiomers, all possible geometric isomers, andpharmaceutically acceptable salts thereof.

The present invention includes all stereoisomers of the compound andpharmaceutically acceptable salts thereof. Further, mixtures ofstereoisomers as well as isolated specific stereoisomers are alsoincluded. During the course of the synthetic procedures used to preparesuch compounds or in using racemization or epimerization proceduresknown to those skilled in the art, the products of such procedures canbe a mixture of stereoisomers.

The term “stereoisomer” as used in the present invention refers to anisomer in which atoms or groups of atoms in the molecule are connectedto each other in the same order but differ in spatial arrangement,including conformational isomers and conformational isomers. Theconfiguration isomers include geometric isomers and optical isomers, andoptical isomers mainly include enantiomers and diastereomers. Theinvention includes all possible stereoisomers of the compound.

Certain of the compounds provided herein may exist as atropisomers,which are conformational stereoisomers that occur when rotation about asingle bond in the molecule is prevented, or greatly slowed, as a resultof steric interactions with other parts of the molecule. The compoundsprovided herein include all atropisomers, both as pure individualatropisomer preparations, enriched preparations of each, or anon-specific mixture of each. Where the rotational barrier about thesingle bond is high enough, and interconversion between conformations isslow enough, separation and isolation of the isomeric species may bepermitted.

The present invention is intended to include all isotopes of atomsoccurring in the present compounds. Isotopes include those atoms havingthe same atomic number but different mass numbers. By way of generalexample and without limitation, isotopes of hydrogen include deuteriumand tritium. The isotopes of hydrogen can be denoted as ¹H (hydrogen),²H (deuterium) and ³H (tritium). They are also commonly denoted as D fordeuterium and T for tritium. In the application, CD₃ denotes a methylgroup wherein all of the hydrogen atoms are deuterium. Isotopes ofcarbon include ¹³C and ¹⁴C. Isotopically-labeled compounds of theinvention can generally be prepared by conventional techniques known tothose skilled in the art or by processes analogous to those describedherein, using an appropriate isotopically-labeled reagent in place ofthe non-labeled reagent.

When a tautomer of the compound exists, the present invention includesany possible tautomers and pharmaceutically acceptable salts thereof,and mixtures thereof, except where specifically stated otherwise.

The pharmaceutical compositions of the present invention comprise thecompound (or the stereoisomer, the astropisomer thereof, thepharmaceutically acceptable salt thereof, the pharmaceuticallyacceptable salt of the stereoisomer, or the pharmaceutically acceptablesalt of the astropisomer) as an active ingredient, and apharmaceutically acceptable carrier and optionally other adjuvants. Thecompositions include compositions suitable for oral, rectal, topical,and parenteral (including subcutaneous, intramuscular, and intravenous)administration, although the most suitable route in any given case willdepend on the particular host, and nature and severity of the conditionsfor which the active ingredient is being administered. Thepharmaceutical compositions may be conveniently presented in unit dosageform and prepared by any of the methods well known in the art ofpharmacy.

In practice, the compounds or a prodrug or a metabolite orpharmaceutically acceptable salts thereof, of this invention can becombined as the active ingredient in intimate admixture with apharmaceutical carrier according to conventional pharmaceuticalcompounding techniques. The carrier may take a wide variety of formsdepending on the form of preparation desired for administration, e.g.oral or parenteral (including intravenous). Thus, the pharmaceuticalcompositions of the present invention can be presented as discrete unitssuitable for oral administration such as capsules, cachets or tabletseach containing a predetermined amount of the active ingredient.Further, the compositions can be presented as a powder, as granules, asa solution, as a suspension in an aqueous liquid, as a non-aqueousliquid, as an oil-in-water emulsion or as a water-in-oil liquidemulsion. In addition to the common dosage forms set out above, thecompound or a pharmaceutically acceptable salt thereof, may also beadministered by controlled release means and/or delivery devices. Thecompositions may be prepared by any of the methods of pharmacy. Ingeneral, such methods include a step of bringing into association theactive ingredient with the carrier that constitutes one or morenecessary ingredients. In general, the compositions are prepared byuniformly and intimately admixing the active ingredient with liquidcarriers or finely divided solid carriers or both. The product can thenbe conveniently shaped into the desired presentation.

The pharmaceutical carrier employed can be, for example, a solid, liquidor gas. Examples of solid carriers include lactose, terra alba, sucrose,talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearicacid. Examples of liquid carriers are sugar syrup, peanut oil, oliveoil, and water. Examples of gaseous carriers include carbon dioxide andnitrogen. In preparing the compositions for oral dosage form, anyconvenient pharmaceutical media may be employed. For example, water,glycols, oils, alcohols, flavoring agents, preservatives, coloringagents, and the like may be used to form oral liquid preparations suchas suspensions, elixirs and solutions; while carriers such as starches,sugars, microcrystalline cellulose, diluents, granulating agents,lubricants, binders, disintegrating agents, and the like may be used toform oral solid preparations such as powders, capsules and tablets.Because of their ease of administration, tablets and capsules are thepreferred oral dosage units whereby solid pharmaceutical carriers areemployed. Optionally, tablets may be coated by standard aqueous ornonaqueous techniques.

A tablet containing the composition of this invention may be prepared bycompression or molding, optionally with one or more accessoryingredients or adjuvants. Compressed tablets may be prepared bycompressing, in a suitable machine, the active ingredient in afree-flowing form such as powder or granules, optionally mixed with abinder, lubricant, inert diluent, surface active or dispersing agent.Molded tablets may be made by molding in a suitable machine, a mixtureof the powdered compound moistened with an inert liquid diluent. Eachtablet preferably contains from about 0.05 mg to about 5 g of the activeingredient and each cachet or capsule preferably containing from about0.05 mg to about 5 g of the active ingredient. For example, aformulation intended for the oral administration to humans may containfrom about 0.5 mg to about 5 g of active agent, compounded with anappropriate and convenient amount of carrier material which may varyfrom about 0.05 to about 95 percent of the total composition. Unitdosage forms will generally contain between from about 0.01 mg to about2 g of the active ingredient, typically 0.01 mg, 0.02 mg, 1 mg, 2 mg, 3mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 25 mg, 50 mg, 100 mg, 200mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, 1000 mg, 1500 mg or 2000 mg.

Pharmaceutical compositions of the present invention suitable forparenteral administration may be prepared as solutions or suspensions ofthe active compounds in water. A suitable surfactant can be includedsuch as, for example, hydroxypropylcellulose. Dispersions can also beprepared in glycerol, liquid polyethylene glycols, and mixtures thereofin oils. Further, a preservative can be included to prevent thedetrimental growth of microorganisms.

Pharmaceutical compositions of the present invention suitable forinjectable use include sterile aqueous solutions or dispersions.Furthermore, the compositions can be in the form of sterile powders forthe extemporaneous preparation of such sterile injectable solutions ordispersions. In all cases, the final injectable form must be sterile andmust be effectively fluid for easy syringability. The pharmaceuticalcompositions must be stable under the conditions of manufacture andstorage; thus, preferably should be preserved against the contaminatingaction of microorganisms such as bacteria and fungi. The carrier can bea solvent or dispersion medium containing, for example, water, ethanol,polyol (e.g., glycerol, propylene glycol and liquid polyethyleneglycol), vegetable oils, and suitable mixtures thereof.

Pharmaceutical compositions of the present invention can be in a formsuitable for topical use such as, for example, an aerosol, cream,ointment, lotion, dusting powder or the like. Further, the compositionscan be in a form suitable for use in transdermal devices. Theseformulations may be prepared, utilizing a compound of this invention ora pharmaceutically acceptable salt thereof, via conventional processingmethods. As an example, a cream or ointment is prepared by admixinghydrophilic material and water, together with about 0.05 wt % to about10 wt % of the compound, to produce a cream or ointment having a desiredconsistency.

Pharmaceutical compositions of this invention can be in a form suitablefor rectal administration wherein the carrier is a solid. It ispreferable that the mixture forms unit dose suppositories. Suitablecarriers include cocoa butter and other materials commonly used in theart. The suppositories may be conveniently formed by first admixing thecomposition with the softened or melted carrier(s) followed by chillingand shaping in molds.

In addition to the aforementioned carrier ingredients, thepharmaceutical formulations described above may include, as appropriate,one or more additional carrier ingredients such as diluents, buffers,flavoring agents, binders, surface-active agents, thickeners,lubricants, preservatives (including antioxidants) and the like.Furthermore, other adjuvants can be included to render the formulationisotonic with the blood of the intended recipient. Compositionscontaining a compound or pharmaceutically acceptable salts thereof, mayalso be prepared in powder or liquid concentrate form.

Generally, dosage levels on the order of from about 0.001 mg/kg to about150 mg/kg of body weight per day are useful in the treatment of theabove-indicated conditions or alternatively about 0.05 mg to about 7 gper patient per day. For example, inflammation, cancer, psoriasis,allergy/asthma, disease and conditions of the immune system, disease andconditions of the central nervous system (CNS), may be effectivelytreated by the administration of from about 0.001 to 50 mg of thecompound per kilogram of body weight per day or alternatively about 0.05mg to about 3.5 g per patient per day.

It is understood, however, that the specific dose level for anyparticular patient will depend upon a variety of factors including theage, body weight, general health, sex, diet, time of administration,route of administration, rate of excretion, drug combination and theseverity of the particular disease undergoing therapy.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is the ball and stick model of the absolute configuration ofCompound 1-2;

FIG. 2 is the ellipsoid model of the absolute configuration of Compound1-2;

FIG. 3 is the graph of unit cell structure of single crystal diffractionfor Compound 1-2;

FIG. 4 is the graph of hydrogen bond of single crystal diffraction forCompound 1-2;

FIG. 5 is the graph of 3D structure-a direction of single crystaldiffraction for Compound 1-2;

FIG. 6 is the graph of 3D structure-b direction of single crystaldiffraction for Compound 1-2;

FIG. 7 is the graph of 3D structure-c direction of single crystaldiffraction for Compound 1-2.

FIG. 8 shows the efficacy of Compound 1-2 and Compound 12-2 in NCI-H1373xenograft model.

FIG. 9 shows the safety of Compound 1-2 in MIA-PaCa-2 model.

METHODS OF PREPARATION

Compounds of the present invention can be synthesized from commerciallyavailable reagents using the synthetic methods described herein. Theexamples which outline specific synthetic route below are meant toprovide guidance to the ordinarily skilled synthetic chemist, who willreadily appreciate that the solvent, concentration, reagent, protectinggroup, order of synthetic steps, time, temperature, and the like can bemodified as necessary, well within the skill and judgment of theordinarily skilled artisan.

The following Examples are provided to better illustrate the presentinvention. All parts and percentages are by weight and all temperaturesare degrees Celsius, unless explicitly stated otherwise. The followingabbreviations in Table 1 have been used in the examples:

TABLE 1 MeOH Methanol EtOH Ethanol DCM Dichloromethane TEA TriethylamineTFA Trifluoroacetic acid DMF N,N-Dimethylformamide DMAN,N-dimethylacetamide THF Tetrahydrofuran MeCN/ACN Acetonitrile HATU2-(7-Azabenzotriazol-1-yl)-N,N,N′,N′- tetramethyluroniumhexafluorophosphate EDCI 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimidehydrochloride HOBT 1-Hydroxybenzotriazole LiHMDS LithiumHexamethyldisilazide Hunig's N,N-Diisopropylethylamine base/DIEA/DIPEAEA Ethyl acetate min Minute(s) h Hour(s) Pre-TLC Preparative thin layerchromatography prep-HPLC Preparative High Performance LiquidChromatography SFC Supercritical fluid chromatography Pd(dppf)Cl₂[1,1′-Bis(diphenylphosphino) ferrocene]dichloropalladium(II) R.T./r.t.Room temperature(20° C.~30° C.) AcOH Acetic acid Pd(PPh₃)₄Tetrakis(triphenylphosphine)palladium NCS N-Chlorosuccinimide Hexn-Hexane PPTS Pyridinium 4-toluenesulfonate IPA Isopropanol DHP3,4-Dihydro-2H-pyran a.q./aq Aqueous AcOK/KOAc Potassium acetate NMPMethyl-2-pyrrolidinone

Example 14-(4-acryloylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 1”)(P)-4-(4-acryloylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-mcthylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 1-1”); and(M)-4-(4-acryloylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 1-2”)

Step 1. 4-methyl-2-(prop-1-en-2-yl)pyridin-3-amine

Into a 350-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed 2-bromo-4-methylpyridin-3-amine (BD, APL099)(15.01 g, 80.25 mmol),4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (BD, AQA827)(13.62 g, 81.05 mmol), Pd(dppf)Cl₂ (5.95 g, 8.03 mmol), K₂CO₃ (33.52 g,240 mmol), dioxane (150 mL) and water (20 mL). The reaction mixture wasstirred at 100° C. for 8 h. The reaction mixture was filtered andconcentrated under vacuum. The residue was applied onto a silica gelcolumn eluted with EA/hexane (v:v=2:3). This resulted in 11.2 g (94%) of4-methyl-2-(prop-1-en-2-yl)pyridin-3-amine as yellow oil. LCMS: m/z=149[M+1]⁺.

Step 2. 2-isopropyl-4-methylpyridin-3-amine (Intermediate A)

Into a 250-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed4-methyl-2-(prop-1-en-2-yl)pyridin-3-amine (11.2 g, 75.67 mmol) and MeOH(100 mL). Palladium on carbon (2.81 g) was added in three portions. Themixture was degassed under vacuum and then purged with H₂ (gas) forthree cycles. The mixture was stirred for 3 h at 25° C. The resultingmixture was filtered, the filtrate was concentrated under vacuum. Thisresulted in 11 g (crude) of 2-isopropyl-4-methylpyridin-3-amine whichwas used directly in the next step. LCMS: m/z=151 [M+1]⁺.

Step 3. 2-cyano-N-(2-isopropyl-4-methylpyridin-3-yl)acetamide

Into a 100-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed 2-cyanoacetic acid (3 g, 35.27 mmol)and DCM (40 mL). Oxalyl chloride (6.2 g, 48.85 mmol) was added indropwise. After the addition, DMF (0.1 mL) was added. The mixture wasstirred for 3 h at 25° C. The resulting solution was concentrated undervacuum. This resulted in 3.10 g (crude) of 2-cyanoacetylchloride whichwas used directly in the next step.

Into a 100-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed 2-isopropyl-4-methylpyridin-3-amine(2.00 g, 13.31 mmol), TEA (5.40 g, 53.36 mmol) and DCM (40 mL) andstirred. The mixture was cooled to 0° C. and then 2-cyanoacetylchloride(3.10 g, crude) was added in dropwise. The resulting solution wasstirred for further 2 h at room temperature. The reaction was thenquenched by the addition of water (100 mL). The resulting solution wasextracted with dichloromethane (3×50 mL), the organic layers werecombined and washed with brine (50 mL), dried over anhydrous Na₂SO₄ andfiltered, the filtrate was concentrated under vacuum and applied onto asilica gel column eluted with EA/hexane (v:v=3:2). This resulted in 1.00g (34%) of 2-cyano-N-(2-isopropyl-4-methyl pyridin-3-yl)acetamide asyellow solid. LCMS: m/z=218 [M+1]⁺.

Step 4.2-cyano-N-(2-isopropyl-4-methylpyridin-3-yl)-3-oxo-3-(2,5,6-trichloropyridin-3-yl)propanamide

Into a 100-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed 2,5,6-trichloronicotinic acid (5.01g, 22.12 mmol) and SOCl₂ (30 mL). The mixture was heated to 80° C. andstirred for 2 h. The solution was concentrated under vacuum. Thisresulted in 5.10 g (crude) of 2,5,6-trichloronicotinoyl chloride whichwas used directly in the next step.

Into a 250-mL round-bottom flask, was placed2-cyano-N-(2-isopropyl-4-methylpyridin-3-yl)acetamide (3.01 g, 13.85mmol) and THF (40 mL). The mixture was stirred at 0° C. NaH (1.16 g,28.99 mmol) was added in three batches. The mixture was stirred at 0° C.for further 40 min. Then 2,5,6-trichloronicotinoyl chloride (3.19 g,13.03 mmol) in THF (10 mL) was added in dropwise. The reaction mixturewas stirred at 25° C. for 2 h. The reaction mixture was concentratedunder vacuum. The resulting crude product was further purified by C₁₈column eluted with ACN/H₂O (v/v=1/3). This resulted in 5.89 g (crude) of2-cyano-N-(2-isopropyl-4-methylpyridin-3-yl)-3-oxo-3-(2,5,6-trichloropyridin-3-yl)propanamideas yellow solid. LCMS: m/z=425 [M+1]+.

Step 5.6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(Intermediate B)

Into a 250-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed2-cyano-N-(2-isopropyl-4-methylpyridin-3-yl)-3-oxo-3-(2,5,6-trichloropyridin-3-yl)propanamide(5.89 g, 13.83 mmol) and THF (70 mL) and stirred at room temperature.NaH (2.73 g, 68.25 mmol) was added in batch-wise. The mixture wasstirred at 50° C. for 2 h. The reaction mixture was concentrated undervacuum. The residue was dissolved in 100 mL water and adjusted pH to 7with AcOH. The resulting solid was filtered and dried under vacuum toprovide 5.85 g (108% in two steps) of6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile as yellow solid. LCMS: m/z=389 [M+1]⁺.

¹H NMR (400 MHz, DMSO-d₆) δ 8.77 (d, J=5.7 Hz, 1H), 8.38 (s, 1H), 7.89(d, J=5.4 Hz, 1H), 3.01-2.88 (m, 1H), 2.19 (s, 3H), 1.21 (d, J=6.9 Hz,3H), 1.14 (d, J=6.9 Hz, 3H).

The mixture of6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileatropisomers (10.59 g, “Intermediate B”) was purified byChiral-Prep-HPLC with the following conditions: Column, CHIRALPAK IC,3.0×100 mm, 3 μm; mobile phase, IPA/ACN=(v/v=1/1); detection wavelength,UV 210 nm. This resulted in 4.99 g (47%) of6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile (the first eluting isomer, “Intermediate B-1”, M or Patropisomer) as a brown solid;

¹H NMR (400 MHz, CD₃OD) δ 8.48 (s, 1H), 8.44 (d, J=5.0 Hz, 1H),7.33-7.22 (m, 1H), 2.76-2.61 (m, 1H), 2.03 (s, 3H), 1.19 (d, J=6.8 Hz,3H), 1.06 (d, J=6.8 Hz, 3H).

And 4.60 g (43%) of6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the second eluting isomer, “Intermediate B-2”, P or M atropisomer) as abrown solid;

¹H NMR (400 MHz, CD₃OD) δ 8.49 (s, 1H), 8.44 (d, J=5.0 Hz, 1H), 7.28 (d,J=5.0 Hz, 1H), 2.76-2.63 (m, 1H), 2.03 (s, 3H), 1.19 (d, J=6.8 Hz, 3H),1.08-1.00 (m, 3H).

Step 6.4,6,7-trichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile (Intermediate B, 980 mg, 2.51 mmol), POCl₃ (1150 mg, 7.50mmol), DIEA (1.32 g, 10.21 mmol) and acetonitrile (12 mL). The mixturewas stirred at 80° C. for 2 h. The reaction mixture was cooled to roomtemperature and concentrated under vacuum. This resulted in crude4,6,7-trichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile whichwas used directly in the next step.

Step 7. tert-butyl4-(6,7-dichloro-3-cyano-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazine-1-carboxylate

Into a 100-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed4,6,7-trichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(1.20 g, crude) and acetonitrile (20 mL). DIEA (660 mg, 5.10 mmol) andtert-butyl piperazine-1-carboxylate (0.57 g, 3.06 mmol) were added. Thereaction mixture was stirred for 2 h at room temperature. The reactionwas then quenched by the addition of water (50 mL). The resultingsolution was extracted with ethyl acetate (3×50 mL), the organic layerswere combined and washed with brine (50 mL), dried over anhydrousNa₂SO₄, filtered and concentrated under vacuum. The residue was purifiedby silica gel column eluted with EA/hexane (v/v=30%˜70%). This resultedin 0.92 g (65% in two steps) of tert-butyl4-(6,7-dichloro-3-cyano-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazine-1-carboxylateas yellow solid. LCMS: m/z=557 [M+1]⁺.

Step 8.4-(4-acryloylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(Intermediate C)

Into a 20-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed tert-butyl4-(6,7-dichloro-3-cyano-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazine-1-carboxylate(920 mg, 1.65 mmol), TFA (4 ml) and DCM (15 mL). The reaction mixturewas stirred at room temperature for 2 h. The reaction mixture wasconcentrated under vacuum. The residue was dissolved by DCM (15 mL) in50-mL round-bottom flask. DIEA (1.02 g, 10.08 mmol) was added. Thereaction mixture was cooled to 0° C. and acryloyl chloride (190 mg, 2.09mmol) was added. The mixture was stirred at room temperature for 2 h.The reaction was then quenched by the addition of water (30 mL). Theresulting solution was extracted with ethyl acetate (3×50 mL), theorganic layers were combined and washed with brine (30 mL), dried overanhydrous Na₂SO₄, filtered and concentrated under vacuum. The residuewas purified by silica gel column eluted with EA/hexane (v/v=40%˜80%).This resulted in 0.86 g (crude) of4-(4-acryloylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileas yellow solid. LCMS: m/z=511 [M+1]⁺.

Step 9. 2-bromo-3,4,5,6-tetrafluoroaniline

Into a 50-mL round-bottom flask was placed 2,3,4,5-tetrafluoroaniline(1.99 g, 12.05 mmol), Sodium acetate (1.32 g, 16.09 mmol), iron (0.10 g,1.79 mmol) and AcOH (7 mL). The reaction mixture was heated to 45° C.This was followed by the added of bromine (3.02 g, 18.90 mmol) in AcOH(7 mL). The reaction mixture was heated to 60° C. and stirred for 1.5 h.The reaction mixture was quenched by the addition of saturated aqueousNa₂S2O₃ (30 mL). The resulting solution was extracted with ethyl acetate(2×50 mL), the organic layers were combined and washed with saturatedaqueous Na₂CO₃ (50 mL) and brine (50 mL), dried over anhydrous Na₂SO₄,filtered and concentrated under vacuum. This resulted in 2.28 g (77%) of2-bromo-3,4,5,6-tetrafluoroaniline as yellow solid. LCMS: m/z=244,246[M+1]⁺.

Step 10.2,3,4,5-tetrafluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline

Into a 150-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed 2-bromo-3,4,5,6-tetrafluoroaniline (2.28 g, 9.35mmol), bis(pinacolato)diboron (3.96 g, 15.59 mmol), Pd(dppf)Cl₂ (1.21 g,1.65 mmol), AcOK (1.84 g, 18.75 mmol) and dioxane (20 mL). The reactionmixture was stirred at 100° C. for 2 h. The reaction mixture wasfiltered and concentrated under vacuum. The residue was applied onto asilica gel column eluted with EA/hexane (v/v=0%-20%). This resulted in2.28 g (81% yield) of2,3,4,5-tetrafluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)anilineas white solid. LCMS: m/z=210 [M+1]⁺.

Step 11.4-(4-acryloylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 1”)

Into a 20-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed4-(4-acryloylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(221 mg, 0.43 mmol),2,3,4,5-tetrafluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline(513 mg, 1.76 mmol), Pd(PPh₃)₄ (151 mg, 0.13 mmol), Na₂CO₃ (161 mg, 1.52mmol), dioxane (5 mL) and water (0.5 mL). The reaction mixture wasstirred at 80° C. for 1 h. The reaction mixture was filtered andconcentrated under vacuum. The resulting crude product was furtherpurified by C₁₈ column eluted with CH₃CN/H₂O (v/v=40%˜80%). Thisresulted in 7 mg (2% yield) of4-(4-acryloylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 1”) as yellow solid. LCMS: m/z=640 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.55 (s, 1H), 8.43 (d, J=5.0 Hz, 1H),7.33-7.20 (m, 1H), 6.87 (dd, J=16.7, 10.6 Hz, 1H), 6.31 (d, J=16.9 Hz,1H), 5.84 (d, J=10.6 Hz, 1H), 3.98 (d, J=23.5 Hz, 8H), 2.73 (d, J=28.0Hz, 1H), 2.00 (d, J=31.9 Hz, 3H), 1.32-1.10 (m, 3H), 1.00 (dd, J=40.7,6.8 Hz, 3H).

The mixture of4-(4-acryloylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileatropisomers (356 mg, several batches) was purified by Chiral-Prep-HPLCwith the following conditions: Column, CHIRALPAK IG, 3 cm×25 cm, 5 um;mobile phase, CO₂:EtOH=55:45; Detection wavelength, UV 220 nm.

This resulted in 175 mg (49.16%) of4-(4-acryloylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the first eluting isomer, “Compound 1-1”) as a yellow solid. LCMS:m/z=640 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.45 (d, J=1.9 Hz, 1H), 8.34 (d, J=5.0 Hz,1H), 7.17 (d, J=5.0 Hz, 1H), 6.78 (dd, J=16.8, 10.6 Hz, 1H), 6.21 (dd,J=16.8, 2.0 Hz, 1H), 5.74 (dd, J=10.6, 2.0 Hz, 1H), 4.02-3.72 (m, 8H),2.70-2.58 (m, 1H), 1.97-1.88 (m, 3H), 1.07 (dd, J=8.2, 6.7 Hz, 3H),0.97-0.88 (m, 3H).

And 184 mg (51.69%) of4-(4-acryloylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the second eluting isomer, “Compound 1-2”) as a yellow solid. LCMS:m/z=640 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.57 (s, 1H), 8.45 (d, J=5.0 Hz, 1H), 7.31 (d,J=4.7 Hz, 1H), 6.88 (dd, J=16.7, 10.7 Hz, 1H), 6.32 (d, J=16.8 Hz, 1H),5.88 (d, J=10.5 Hz, 1H), 4.08-3.92 (m, 8H), 2.81-2.66 (m, 1H), 2.06-1.98(m, 3H), 1.18 (t, J=7.4 Hz, 3H), 1.06-0.96 (in, 3H).

About 10 mg Compound 1-2 was taken into a glass vial and dissolved with0.8 mL ethanol and 0.4 mL n-heptane. The clear solution was evaporatedto dryness at room temperature through a small hole to obtain thebulk-like crystals as the sample of the testing of single crystaldiffraction using the following instrument and parameters in Table 2:

TABLE 2 Instrument and Parameters Instrument Single CrystalDiffractometer Model Bruker SMART APEX II Detector Model 4K CCD SourcesEnhance Cu radiation Lens Temperature 293.44 K Wavelength  1.54 Å

The results are shown in Table 3, Table 4, Table 5, Table 6 and Table 7.

Crystallographic Data

TABLE 3 Crystallographic Data and Structure Refinement for Compound 1-2Phase Data Formula C₃₁H₂₆ClF₄N₇O₂ Formula Weight 640.04 Crystal SystemMonoclinic Space group C2 Cell Parameters a = 20.6796 (7) Å; b = 11.2352(4) Å; c = 14.0148 (5) Å; α = γ = 90.00°; β = 91.971 (2) Cell Ratio a/b= 1.8406; b/c = 0.8017; c/a = 0.6777 Z 4 Cell Volume 3254.3 (2) Å³ Calc.density 1.306 g/cm³ Flack 0.07 (3) R-indices R₁ 0.0962 R-indices WR₂0.1665 Goodness-of-Fit, S 1.015 R_(sigma) 0.0799 R_(int) 0.0923

Molecular Structure of Compound 1-2

TABLE 4 Molecular Structure of Compound 1-2 Results Molecular AbsoluteConfiguration in single crystal Planar chirality (Axis chirality) MMolecular Ball Figure 1 and Stick Model in single crystal MolecularEllipsoid Figure 2 Model in single crystal Structure of AbsoluteConfiguration

Unit cell Structure Figure 3 Main Hydrogen Figure 4 Bond In singlecrystal Proton Transfer in — Single crystal 3D Structure-a NetworkDirection Looking From a Figure 5 Direction 3D Structure-b Chained andLaminated Direction Looking From b Figure 6 3D Structure-c Chained andLaminated Direction Looking From c Figure 7 Direction

Crystal Structure Solution Data

TABLE 5 Atomic Coordinates (×10⁴) and Equivalent Isotropic DisplacementParameters (Å² × 10³) for Compound 1-2. U(eq) is Defined as One Third ofthe Trace of the Orthogonalized U^(ij) Tensor. Atom x y z U(eq) C116246.0(6) 1711.4(11) 4631.9(9)  90.2(4) F1 4785.8(15)   2906(3)  3954(3) 117.0(11) F2 4111.9(15)   1499(4)   2704(3) 136.7(14) F3  4659(2)   862(4)   1040(3) 148.9(16) F4   5807(2)   1798(5)    582(2)153.1(16) O1 6947.1(19)   8554(3)   4282(3) 103.3(12) O2   6947(2)  3659(5)   9789(3) 127.4(16) N1 6087.9(17)   4937(3)   3501(3)  68.6(9)N2 6491.6(17)   6770(4)   3932(2)  72.5(9) N3   7296(3)   8781(6)  6695(4)   133(2) N4 6971.7(18)   5546(4)   6708(3)  76.7(10) N6  6493(2)   3262(5)   1777(3) 105.7(15) N7   5536(3)   8178(6)   1965(5)  148(2) C1   5043(3)   2606(5)   3116(4)  84.7(13) C2   4701(3)  1929(5)   2488(5)  93.3(16) C3   4966(3)   1641(6)   1637(4) 100.9(17)C4   5555(3)   2081(5)   1423(4)  97.7(17) C5   5901(2)   2829(4)  2034(3)  79.3(13) C6   5646(2)   3082(4)   2919(3)  71.5(11) C7  6001(2)   3774(4)   3662(3)  70.8(12) C8   6264(2)   3232(4)   4494(3) 69.3(11) C9   6532(2)   3936(4)   5196(3)  70.3(11) C10   6571(2)  5161(4)   5077(3)  64.6(10) C11   6846(2)   5977(5)   5801(3) 75.2(13) C12   6960(2)   7121(4)   5505(3)  73.5(12) C13   6820(2)  7551(5)   4547(4)  79.2(13) C14   6379(2)   5597(4)   4184(3) 68.9(11) C15   7166(3)   8039(6)   6156(4)  94.6(16) C16   6309(3)  7206(4)   2980(4)  86.0(15) C17   6749(4)   7054(5)   2262(4)   110(2)C18   6535(6)   7460(8)   1367(5)   146(3) C19   5946(6)   8011(11)  1271(7)   166(4) C20   5716(3)   7763(6)   2858(4)  98.4(17) C21  5247(3)   7954(6)   3613(5)   114(2) C23   4722(4)   7012(9)   3532(8)  173(4) C22   4967(6)   9207(9)   3602(9)   199(4) C24   7389(4)  6458(7)   2421(6)   142(3) C25   6453(5)   5039(9)   7253(8)   79(2)C26   6758(4)   4128(9)   7917(7)   85(2) N5   7274(4)   4605(7)  8506(6)  85.7(18) C27   7753(4)   5199(8)   7924(6)  83.4(19) C28  7476(6)   6148(10)   7310(8)   84(2) C25′   6610(9)   4538(16)  7194(14)   85(3) C26′   6480(7)   4955(14)   8202(9)   87(2) N5′  7092(6)   5244(12)   8726(9)  88.7(18) C27′   7501(7)   6051(13)  8300(9)   88(2) C28′   7620(9)   5650(16)   7238(14)   87(3) C29  7324(3)   4417(7)   9488(4) 109.7(19) C30   7861(3)   4907(7) 10040(4)   112(2) C31   8076(4)   4358(8)  10783(5)   128(2)

TABLE 6 Hydrogen Coordinates (×10⁴) and Isotropic DisplacementParameters (Å² × 10³) for Compound 1-2. Atom x y z U(eq) H6A 6646 30671236 127 H6B 6709 3728 2156 127 H9 6691 3591 5761 84 H18 6790 7358 839175 H19 5823 8293 668 199 H21 5478 7841 4229 137 H23A 4551 6982 2887 259H23B 4381 7207 3953 259 H23C 4901 6250 3705 259 H22A 4702 9317 3034 299H22B 5314 9777 3614 299 H22C 4710 9319 4153 299 H24A 7549 6613 3059 214H24B 7689 6762 1973 214 H24C 7339 5615 2331 214 H25A 6133 4666 6828 95H25B 6242 5654 7614 95 H26A 6429 3808 8321 102 H26B 6924 3477 7542 102H27A 7955 4610 7525 100 H27B 8088 5538 8342 100 H28A 7804 6496 6919 101H28B 7288 6770 7692 101 H25C 6206 4369 6849 102 H25D 6870 3820 7212 102H26C 6205 5655 8174 104 H26D 6255 4335 8539 104 H27C 7910 6088 8659 106H27D 7307 6838 8301 106 H28C 7888 6231 6926 104 H28D 7842 4889 7236 104H30 8050 5618 9856 135 H31A 7886 3647 10965 153 H31B 8423 4669 11143 153

TABLE 7 Anisotropic Displacement Parameters (Å² × 10³) for Compound 1-2.The Anisotropic Displacement Factor Exponent Takes the Form: −2π²[ h²a*²U₁₁ + ... + 2 h k a* b* U₁₂] Atom U₁₁ U₂₂ U₃₃ U₂₃ U₁₃ U₁₂ C11 104.2(9)   71.6(7)   94.8(9)   11.9(6)   4.1(7)   −1.6(7) F1 91(2)142(3)  120(3)  −12(2)    35.8(19)   −15.5(19) F2 93(2) 156(3)  159(3) 23(3) −15(2)  −45(2) F3 178(3)  164(3)  101(3)  13(2) −45(2)  −73(3) F4175(3)  205(4)  80(2) −41(3)  9(2) −65(3) O1 116(3)  81(2) 113(3)  11(2)−1(2)  −27(2) O2 126(3)  181(5)  76(3) 27(3) 11(2)  −32(3) N1 72(2)63(2) 71(2)    3.0(17)   4.7(19)    −2.8(17) N2 83(2) 75(2) 60(2)  12.0(19)   3.0(18) −11(2) N3 137(5)  136(5)  125(5)  −46(4)  7(4)−27(4) N4 75(2) 96(3) 59(2)    2.3(18)   5.7(19) −17(2) N6 97(3) 155(4) 66(3) −17(3)  19(2)  −41(3) N7 140(5)  166(6)  137(5)  70(5) −27(4) −38(4) C1 80(3) 90(3) 85(4)  2(3) 12(3)   −4(3) C2 72(3) 93(4) 113(4) 19(3) −14(3)  −25(3) C3 107(4)  109(4)  85(4) 15(3) −33(3)  −27(4) C4109(4)  128(5)  55(3) −7(3) −6(3)  −19(3) C5 89(3) 87(3) 62(3)  5(2)0(3) −11(3) C6 75(3) 73(3) 66(3) −1(2) 5(2) −12(2) C7 68(3) 76(3) 70(3) 2(2) 21(2)   0(2) C8 77(3) 60(3) 71(3) 15(2) 7(2)  −1(2) C9 68(3) 80(3)62(3)  2(2) 9(2)  −2(2) C10 68(3) 68(3) 58(2)    9.1(18) 5(2)  −3(2) C1164(3) 98(4) 64(3) −4(2) 11(2)  −12(2) C12 68(3) 84(3) 70(3) −9(2) 9(2)−18(2) C13 73(3) 83(3) 82(3)  3(3) 6(2) −15(3) C14 69(3) 65(3) 73(3) 2(2) 20(2)  −10(2) C15 95(4) 102(4)  88(4) −12(3)  15(3)  −10(3) C16111(4)  78(3) 69(3) 15(2) 1(3) −22(3) C17 155(6)  101(4)  76(4) 20(3)29(4)  −26(4) C18 219(10) 138(6)  82(5) 28(4) 21(6)  −29(6) C19 186(9) 200(10) 111(7)  70(6) −19(7)  −54(8) C20 95(4) 110(4)  90(4) 35(3)−6(3)  −22(3) C21 85(4) 111(5)  145(6)  29(4) 2(4)  1(3) C23 101(5) 180(9)  239(10) −11(7)  35(6)  −29(5) C22 213(10) 132(7)  252(12) 30(7)−6(9)   37(7) C24 162(6)  114(6)  156(7)   6(4) 67(5)   10(5) C25 72(5)94(5) 71(5) 14(4) 9(4) −14(4) C26 79(5) 95(5) 81(5) 13(4) 7(4) −16(4) N592(4) 91(5) 74(4) 12(3) 8(3)  −7(4) C27 88(5) 97(5) 65(4)  0(4) 0(4)−16(4) C28 87(5) 97(6) 67(5) −2(4) 0(4) −15(4) C25′ 90(6) 93(6) 72(6) 7(5) 12(5)  −12(5) C26′ 92(5) 95(5) 74(5)  7(4) 11(4)   −8(4) N5′ 97(4)95(5) 75(4)  8(4) 7(3)  −9(4) C27′ 97(5) 93(5) 75(5)  8(4) 5(4) −12(4)C28′ 95(6) 91(6) 75(6)  7(5) 3(5) −16(5) C29 107(5)  150(6)  72(4) 10(3)11(3)  −23(4) C30 138(6)  131(5)  68(4)  3(3) −1(4)  −18(4) C31 129(6) 164(6)  90(5) −6(4) 9(4)  −4(5)

The following conclusions were obtained by crystal structure analysis:

The crystal system of Compound 1-2 is monoclinic and has two symmetriesand one centering vector, space group is C2.

The single crystal of Compound 1-2 have four molecules in a unit cell.

Compound 1-2 has one planar chirality (axis chirality). Absoluteconfiguration is M.

Unit cell has two kinds of hydrogen bonds, they are respectivelyO2-H6A-N6, N3-H6B—N6. The three-dimensional structure of the chain,laminate and network is formed through hydrogen bond and van der Waalsforce in the crystal.

Example 24-(4-acryloylpiperazin-1-yl)-7-(3-amino-2,6-dichloro-4,5-difluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 2”)

Step 1.2-(3,4-difluoro-5-nitrophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

Into a 40-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed 5-bromo-1,2-difluoro-3-nitrobenzene (207 mg,0.38 mmol), bis(pinacolato)diboron (2.16 g, 8.51 mmol), Pd(dppf)Cl₂(0.322 g, 0.44 mmol), KOAc (1.199 g, 12.22 mmol) and dioxane (10 mL).The reaction mixture was stirred at 100° C. for 2 h. The reactionmixture was filtered and concentrated under vacuum. The residue wasapplied onto a silica gel column eluted with EA/hexane (v/v=0%-5%). Thisresulted in 1.528 g (crude) of2-(3,4-difluoro-5-nitrophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolaneas off-white solid.

Step 2.2,3-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline

Into a 40-mL round-bottom flask was placed2-(3,4-difluoro-5-nitrophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(1.489 g, 5.22 mmol), iron (1.546 g, 27.68 mmol), NH₄Cl (2.654 g, 49.62mmol), EtOH (15 mL) and H₂O (5 mL). The reaction mixture was stirred at80° C. for 1 h. The reaction mixture was filtered and concentrated undervacuum. The residue was washed with DCM (20 mL), filtered andconcentrated under vacuum. This resulted in 0.957 g of2,3-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline asoff-white solid. LCMS: m/z=256 [M+1]⁺.

Step 3.4-(4-acryloylpiperazin-1-yl)-7-(3-amino-4,5-difluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Into a 20-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed4-(4-acryloylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(0.21 g, 0.41 mmol),2,3-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline(0.279 g, 1.09 mmol), Pd(PPh₃)₄ (57 mg, 0.05 mmol), Na₂CO₃ (206 mg, 1.94mmol), dioxane (5 mL) and water (1 mL). The reaction mixture was stirredat 80° C. for 1 h. The reaction mixture was filtered and concentratedunder vacuum. The residue was applied onto a silica gel column elutedwith EA/hexane (v/v=50%-100%). This resulted in 125 mg of4-(4-acryloylpiperazin-1-yl)-7-(3-amino-4,5-difluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileas yellow solid. LCMS: m/z=604 [M+1]⁺.

Step 4.4-(4-acryloylpiperazin-1-yl)-7-(3-amino-2,6-dichloro-4,5-difluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 2”)

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed4-(4-acryloylpiperazin-1-yl)-7-(3-amino-4,5-difluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(103 mg, 170.52 μmol), NCS (51 mg, 381.93 μmol) and DMA (1.5 mL). Thereaction mixture was stirred for 15 h at room temperature. The reactionwas then quenched by the addition of H₂O (20 mL). The resulting solutionwas extracted with ethyl acetate (3×20 mL), the organic layers werecombined and concentrated under vacuum. The residues was purified byprep-HPLC eluted with CH₃CN/H₂O (v/v=7/1). This resulted in 30 mg (26%)of4-(4-acryloylpiperazin-1-yl)-7-(3-amino-2,6-dichloro-4,5-difluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 2”). LCMS: m/z=672 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.56 (s, 1H), 8.40 (d, J=5.0 Hz, 1H),7.29-7.15 (m, 1H), 6.87 (dd, J=16.8, 10.6 Hz, 1H), 6.30 (d, J=16.8 Hz,1H), 5.83 (d, J=10.7 Hz, 1H), 4.30-3.77 (m, 8H), 2.85-2.66 (m, 1H), 1.99(s, 3H), 1.18 (d, J=6.7 Hz, 3H), 1.01 (d, J=6.8 Hz, 3H).

Example 34-(4-acryloylpiperazin-1-yl)-7-(2-amino-3-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 3”)

Step 1. 2-bromo-3,4,5-trifluoroaniline

Into a 100-mL round-bottom flask was placed2-bromo-3,4,5-trifluoro-1-nitrobenzene (2.64 g, 10.31 mmol), iron (5.71g, 102.25 mmol), ammonium chloride (5.67 g, 106.00 mmol), ethanol (25mL) and water (25 mL). The reaction mixture was heated to 55° C. andstirred for 1.5 h. The reaction mixture was filtered and the filtratewas concentrated under vacuum. The residues was dissolved in ethanol (30mL) and applied onto a C₁₈ column eluted with CH₃CN/H₂O (v:v=9:1). Thisresulted in 1.33 g (57%) of 2-bromo-3,4,5-trifluoroaniline. LCMS:m/z=226, 228 [M+1]⁺.

Step 2.3,4,5-trifluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline

Into a 40-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed 2-bromo-3,4,5-trifluoroaniline (2.99 g, 13.23mmol), bis(pinacolato)diboron (10.20 g, 40.17 mmol), Pd(dppf)Cl₂ (0.98g, 1.34 mmol), KOAc (4.18 g, 42.59 mmol) and dioxane (40 mL). Thereaction mixture was stirred at 100° C. for 5 h. The reaction mixturewas concentrated under vacuum. The residue was applied onto a silica gelcolumn eluted with EA/hexane (v/v=0%-10%). This resulted in 5.34 g(crude) of3,4,5-trifluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline.

Step 3.4-(4-acryloylpiperazin-1-yl)-7-(6-amino-2,3,4-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Into a 20-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed4-(4-acryloylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(0.547 g, 1.08 mmol),3,4,5-trifluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline(0.279 g, 1.09 mmol), Pd(PPh₃)₄ (1.376 g, 5.04 mmol), Na₂CO₃ (0.347 g,3.27 mmol), dioxane (5 mL) and water (1 mL). The reaction mixture wasstirred at 80° C. for 4 h. The reaction mixture was filtered andconcentrated under vacuum. The residue was applied onto a silica gelcolumn eluted with EA/hexane (v/v=50%-100%). This resulted in 115 mg of4-(4-acryloylpiperazin-1-yl)-7-(6-amino-2,3,4-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileas yellow solid. LCMS: m/z=622 [M+1]⁺.

Step 4.4-(4-acryloylpiperazin-1-yl)-7-(2-amino-3-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 3”)

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed4-(4-acryloylpiperazin-1-yl)-7-(6-amino-2,3,4-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(100 mg, 160.76 μmol), NCS (0.035 g, 262.11 μmol) and AcOH (1 mL). Thereaction mixture was stirred for 7 h at 35° C. and 15 h at roomtemperature.

The reaction was then quenched by the addition of saturated aqueousNaHCO₃ (30 mL). The resulting solution was extracted with ethyl acetate(2×10 mL), the organic layers were combined and concentrated undervacuum. The residues was purified by prep-HPLC eluted with CH₃CN/H₂O(v/v=6/1). This resulted in 6 mg (5%) of4-(4-acryloylpiperazin-1-yl)-7-(2-amino-3-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 3”) as yellow solid. LCMS: m/z=656 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.66-8.51 (m, 1H), 8.43 (d, J=5.1 Hz, 1H),7.26 (d, J=5.1 Hz, 1H), 6.88 (dd, J=16.8, 10.7 Hz, 1H), 6.31 (d, J=16.7Hz, 1H), 5.84 (d, J=10.7 Hz, 1H), 4.06-3.92 (m, 8H), 2.84-2.62 (m, 1H),2.06-1.92 (m, 3H), 1.17 (dd, J=9.6, 7.0 Hz, 3H), 1.06-0.94 (m, 3H).

Example 44-(4-acryloylpiperazin-1-yl)-7-(2-amino-3,5-dichloro-4,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 4”)

Step 1.3,5-difluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline

Into a 100-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed 2-bromo-3,5-difluoroaniline (3.01 g, 14.47mmol), bis(pinacolato)diboron (6.46 g, 25.43 mmol), Pd(dppf)Cl₂ (1.03 g,1.41 mmol), KOAc (4.20 g, 42.83 mmol), dioxane (20 mL). The reactionmixture was stirred at 80° C. for 12 h. The reaction mixture wasfiltered and concentrated under vacuum. The residue was purified bysilica gel column eluted with EA/hexane (v/v=1/19). This resulted in6.01 g (crude) of3,5-difluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline asyellow oil. LCMS: m/z=256 [M+1]⁺.

Step 2. tert-butyl4-(7-(2-amino-4,6-difluorophenyl)-6-chloro-3-cyano-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazine-1-carboxylate

Into a 100-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed3,5-difluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline(5.86 g, 23.00 mmol), tert-butyl4-(6,7-dichloro-3-cyano-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazine-1-carboxylate(1.29 g, 2.31 mmol), Pd(PPh₃)₄ (0.73 g, 0.63 mmol), Na₂CO₃ (0.84 g, 7.89mmol), dioxane (15 mL) and water (2 mL). The reaction mixture wasstirred at 80° C. for 2 h. The reaction mixture was filtered andconcentrated under vacuum. The residue was purified by silica gel columneluted with EA/hexane (v/v=1/1). This resulted in 1.42 g (crude) oftert-butyl4-(7-(2-amino-4,6-difluorophenyl)-6-chloro-3-cyano-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazine-1-carboxylateas yellow solid. LCMS: m/z=650 [M+1]⁺.

Step 3. tert-butyl4-(7-(2-amino-3,5-dichloro-4,6-difluorophenyl)-6-chloro-3-cyano-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazine-1-carboxylate

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed tert-butyl4-(7-(2-amino-4,6-difluorophenyl)-6-chloro-3-cyano-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazine-1-carboxylate(1.21 g, 1.87 mmol), NCS (0.49 g, 3.73 mmol) and AcOH (30 mL). Thereaction mixture was stirred for 48 h at room temperature. The reactionwas then quenched by the addition of water (100 mL). The resultingsolution was extracted with ethyl acetate (2×100 mL), the organic layerswere combined and washed with brine (10 mL), dried over anhydrousNa₂SO₄, filtered and concentrated under vacuum. The residue was purifiedby silica gel column eluted with EA/hexane (v/v=1/1). This resulted in0.53 g (crude) of tert-butyl4-(7-(2-amino-3,5-dichloro-4,6-difluorophenyl)-6-chloro-3-cyano-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazine-1-carboxylateas yellow solid. LCMS: m/z=718[M+1]⁺.

Step 4.4-(4-acryloylpiperazin-1-yl)-7-(2-amino-3,5-dichloro-4,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 4”)

Into a 20-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed tert-butyl4-(7-(2-amino-3,5-dichloro-4,6-difluorophenyl)-6-chloro-3-cyano-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazine-1-carboxylate(0.53 g, 0.73 mmol), TFA (5 ml) and DCM (20 mL). The reaction mixturewas stirred at room temperature for 2 h. The reaction mixture wasconcentrated under vacuum. The residue was dissolved by DCM (10 mL) in25-mL round-bottom flask. DIEA (1.22 g, 9.48 mmol) was added. Thereaction mixture was cooled to 0° C. and acryloyl chloride (0.08 g, 0.88mmol) was added. The mixture was stirred at room temperature for 0.5 h.The reaction mixture was filtered and concentrated under vacuum. Theresidue was purified by Prep-HPLC (CH₃CN/H₂O (v/v=7/3)). This resultedin 239 mg (50% in two steps) of4-(4-acryloylpiperazin-1-yl)-7-(2-amino-3,5-dichloro-4,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 4”) as yellow solid. LCMS: m/z=672 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.55 (s, 1H), 8.43 (s, 1H), 7.26 (s, 1H), 6.87(dd, J=16.1, 11.1 Hz, 1H), 6.31 (d, J=16.3 Hz, 1H), 5.84 (d, J=10.1 Hz,1H), 4.25-3.75 (m, 8H), 2.85-2.60 (m, 1H), 2.14-1.88 (m, 3H), 1.25-0.87(m, 6H).

Example 54-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(2-amino-3,5-dichloro-4,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 5”)

Step 1.4-((3R,5S)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(Intermediate D)

Into a 500-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(6.21 g, 15.95 mmol), POCl₃ (6.88 g, 48.87 mmol), DIEA (6.80 g, 52.61mmol) and acetonitrile (100 mL). The mixture was stirred for 2 h at 80°C. The reaction was cooled to room temperature and concentrated undervacuum. This resulted in4,6,7-trichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrilewhich was used directly in the next step.

Into a 500-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed4,6,7-trichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(crude), DIEA (6.80 g, 52.61 mmol) and acetonitrile (100 mL),(2S,6R)-2,6-dimethylpiperazine (2.17 g, 19.00 mmol) was added. Themixture was stirred for 1 h at room temperature. The resulting solutionwas concentrated under vacuum and applied onto a silica gel columneluted with EA/hexane (v/v=2/1). This resulted in 4.30 g (50% yield) of4-((3R,5S)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileas yellow solid. LCMS: m/z=539 [M+1]⁺.

Step 2.3,5-difluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline

Into a 40-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed 2-bromo-3,5-difluoroaniline (2.06 g, 9.91 mmol),bis(pinacolato)diboron (4.95 g, 19.49 mmol), Pd(dppf)Cl₂ (809 mg, 1.11mmol), KOAc (2.23 g, 22.69 mmol), dioxane (20 mL). The reaction mixturewas stirred at 100° C. for 19 h. The reaction mixture was concentratedunder vacuum. The residue was applied onto a C₁₈ column eluted withCH₃CN/H₂O (v/v=5%-100%). This resulted in 1.19 g of3,5-difluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline asoff-white solid. LCMS: m/z=256 [M+1]⁺.

Step 3.4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(2-amino-4,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydroquinoline-3-carbonitrile

Into a 20-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydroquinoline-3-carbonitrile (0.196 g, 0.36 mmol),3,5-difluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline(0.27 g, 1.06 mmol), Pd(PPh₃)₄ (78 mg, 0.07 mmol), Na₂CO₃ (207 mg, 1.95mmol), dioxane (5 mL) and water (1 mL). The reaction mixture was stirredat 80° C. for 1 h. The reaction mixture was filtered and concentratedunder vacuum. The residues was applied onto a silica gel column elutedwith EA/hexane (v/v=50%-100%). This resulted in 223 mg of4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(2-amino-4,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydroquinoline-3-carbonitrileas yellow solid. LCMS: m/z=632 [M+1]⁺.

Step 4.4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(2-amino-3,5-dichloro-4,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydroquinoline-3-carbonitrile(“Compound 5”)

Into a 20-mL round-bottom flask was placed4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(2-amino-4,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydroquinoline-3-carbonitrile(223 mg, 0.35 mmol), NCS (98 mg, 0.73 mmol) and HOAc (3 mL). Thereaction mixture was stirred at room temperature for 1 day. The reactionwas heated to 45° C. and stirred at this temperature for 2 hours. Thereaction mixture was concentrated under vacuum. The residues waspurified by Prep-HPLC CH₃CN/H₂O (0.05% NH₄HCO₃) (v/v=2/1). This resultedin 55 mg (22%) of4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(2-amino-3,5-dichloro-4,6-difluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydroquinoline-3-carbonitrile(“Compound 5”) as yellow solid. LCMS: m/z=700 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.83 (s, 1H), 8.43 (d, J=5.0 Hz, 1H), 7.27 (d,J=5.0 Hz, 1H), 6.95-6.82 (m, 1H), 6.33 (d, J=16.5 Hz, 1H), 5.84 (d,J=10.8 Hz, 1H), 4.77 (s, 2H), 4.13-3.92 (m, 2H), 3.84 (d, J=9.4 Hz, 2H),2.80-2.60 (m, 1H), 2.08-1.93 (m, 3H), 1.72-1.59 (m, 6H), 1.25-1.10 (m,3H), 1.10-0.89 (m, 3H).

Example 64-(4-acryloylpiperazin-1-yl)-7-(3-amino-2,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 6”)

Step 1.4-(4-acryloylpiperazin-1-yl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-7-(tributylstannyl)-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Into a 30-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed4-(4-acryloylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(1.039 g, 2.04 mmol), 1,1,1,2,2,2-Hexabutyl-distannane (4.124 g, 7.11mmol), Pd(PPh₃)₄ (0.566 g, 0.49 mmol) and dioxane (10 mL). The reactionmixture was stirred at 100° C. for 1 d. The reaction mixture wasfiltered and concentrated under vacuum. The residues was purified bysilica gel column eluted with EA/hexane (v/v=2/1). This resulted in 273mg (17%) of4-(4-acryloylpiperazin-1-yl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-7-(tributylstannyl)-1,2-dihydro-1,8-naphthyridine-3-carbonitrileas yellow solid. LCMS: m/z=767 [M+1]⁺.

Step 2.4-(4-acryloylpiperazin-1-yl)-7-(3-amino-2,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 6”)

Into a 25-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed4-(4-acryloylpiperazin-1-yl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-7-(tributylstannyl)-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(0.272 g, 0.36 mmol), 2,3,4,6-tetrafluoro-5-iodoaniline (0.630 g, 2.17mmol), Pd(PPh₃)₄ (0.187 g, 0.16 mmol), cuprous iodide (0.294 g, 1.54mmol), dioxane (10 mL). The reaction mixture was stirred at 100° C. for1 d. The reaction mixture was filtered and concentrated under vacuum.The residues was purified by prep-HPLC eluted with ACN/H₂O (v/v=1/2).This resulted in 6 mg (3%) of4-(4-acryloylpiperazin-1-yl)-7-(3-amino-2,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 6”) as yellow solid. LCMS: m/z=640 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.56 (s, 1H), 8.42 (d, J=4.9 Hz, 1H),7.31-7.20 (m, 1H), 6.87 (dd, J=16.7, 10.6 Hz, 1H), 6.30 (dd, J=16.8, 1.7Hz, 1H), 5.83 (dd, J=10.6, 1.8 Hz, 1H), 4.08-3.87 (m, 8H), 2.79-2.62 (m,1H), 2.00 (s, 3H), 1.17 (d, J=6.8 Hz, 3H), 0.98 (d, J=6.7 Hz, 3H).

Example 74-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 7”)

Step 1.4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(5-amino-2,3,4-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Into a 30-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile (0.822 g, 1.52 mmol),(5-amino-2,3,4-trifluorophenyl) boronic acid (0.608 g, 2.23 mmol),Pd(PPh₃)₄ (0.650 g, 0.56 mmol), Na₂CO₃ (0.575 g, 5.43 mmol), dioxane (10mL) and water (2 mL). The reaction mixture was stirred at 80° C. for 1h. The reaction mixture was filtered and concentrated under vacuum. Theresidue was purified by silica gel column eluted with EA/hexane(v/v=2/1). This resulted in 1.079 g (91%) of4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(5-amino-2,3,4-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileas yellow solid. LCMS: m/z=650 [M+1]⁺.

Step 2.4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 7”)

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(5-amino-2,3,4-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(0.622 g, 0.96 mmol), NCS (0.703 g, 5.27 mmol), and AcOH (5 mL). Themixture was stirred at r.t. for 2 d. The reaction was then quenched bythe addition of water (10 mL). The resulting solution was extracted withethyl acetate (3×10 mL), the organic layers were combined and washedwith brine (1×50 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under vacuum. The residues was purified by Prep-HPLC(CH₃CN/H₂O (v/v=6/4)). This resulted in 35 mg (5%) of4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 7”) as yellow solid. LCMS: m/z=684 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.83 (s, 1H), 8.42 (d, J=5.0 Hz, 1H),7.22-7.10 (m, 1H), 6.88 (dd, J=16.7, 10.7 Hz, 1H), 6.32 (d, J=16.5 Hz,1H), 5.74 (dd, J=10.6, 2.0 Hz, 1H), 4.76 (s, 2H), 4.06-3.73 (m, 4H),2.82-2.69 (m, 1H), 2.10-1.94 (m, 3H), 1.63-1.35 (m, 6H), 1.17 (d, J=6.5Hz, 3H), 1.06-0.89 (m, 3H).

Example 84-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(3-amino-2,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 8”)

Step 1.4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Into a 20-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed6,7-dichloro-4-((3S,5R)-3,5-dimethylpiperazin-1-yl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(6.503 g, 13.39 mmol), DIEA (5.21 g, 40.31 mmol), DCM (6 mL) andacryloyl chloride (1.18 g, 13.04 mmol). The reaction mixture was stirredat room temperature for 0.5 h. The reaction mixture was filtered andconcentrated under vacuum. The residue was purified by silica gel columneluted with EA/hexane (v/v=1/1). This resulted in 1.80 g (crude) of4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileas yellow solid. LCMS: m/z=539 [M+1]⁺.

Step 2.4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-7-(tributylstannyl)-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Into a 20-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile (1.77 g, 3.28 mmol),hexabutyldistannane (23.85 g, 6.64 mmol), Pd(PPh₃)₄ (0.26 g, 0.23 mmol)and dioxane (20 mL). The reaction mixture was stirred at 110° C. for 18h. The reaction mixture was filtered and concentrated under vacuum. Theresidue was purified by silica gel column eluted with EA/hexane(v/v=2/1). This resulted in 0.52 g (crude) of4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-7-(tributylstannyl)-1,2-dihydro-1,8-naphthyridine-3-carbonitrileas yellow solid. LCMS: m/z=795 [M+1]⁺.

Step 3.4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(3-amino-2,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile.(“Compound 8”)

Into a 20-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-7-(tributylstannyl)-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(0.40 g, 0.51 mmol), 2,3,4,6-tetrafluoro-5-iodoaniline (0.27 g, 0.93mmol), Pd(PPh₃)₄ (0.27 g, 0.23 mmol), CuI (0.40 g, 2.11 mmol), DMA (15mL). The reaction mixture was stirred at 90° C. for 18 h. The reactionmixture was filtered and concentrated under vacuum. The residue waspurified by Prep-HPLC (CH₃CN/H₂O (v/v=7/3)). This resulted in 8 mg (2.3%in two steps) of4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(3-amino-2,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 8”) as yellow solid. LCMS: m/z=668 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.73 (s, 1H), 8.34 (d, J=5.0 Hz, 1H),7.23-7.15 (m, 1H), 6.79 (dd, J=16.7, 10.7 Hz, 1H), 6.23 (dd, J=16.7, 2.0Hz, 1H), 5.74 (dd, J=10.6, 2.0 Hz, 1H), 4.73-4.58 (m, 2H), 3.95-3.70 (m,4H), 2.66-2.57 (m, 1H), 1.93 (d, J=8.4 Hz, 3H), 1.55-1.57 (m, 6H), 1.07(d, J=6.8 Hz, 3H), 0.89 (d, J=5.7 Hz, 3H).

Example 9(P)-4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile;or(M)-4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile;(“Compound 9”)

Step 1.6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Into a 250-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed2-cyano-N-(2-isopropyl-4-methylpyridin-3-yl)-3-oxo-3-(2,5,6-trichloropyridin-3-yl)propanamide(5.89 g, 13.83 mmol) and THF (70 mL), the mixture was stirred at roomtemperature. This was followed by the addition of NaH (2.73 g, 68.25mmol) in batch-wise. The mixture was stirred at 50° C. for 2 h. Thereaction mixture was concentrated under vacuum. The residue wasdissolved in 100 mL water and adjusted pH to 7 with AcOH. The resultingsolid was filtered and dried under vacuum to provide 5.85 g (108% in twosteps) of6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileas yellow solid. LCMS: m/z=389 [M+1]⁺.

¹H NMR (400 MHz, DMSO-d₆) δ 8.77 (d, J=5.7 Hz, 1H), 8.38 (s, 1H), 7.89(d, J=5.4 Hz, 1H), 3.01-2.88 (m, 1H), 2.19 (s, 3H), 1.21 (d, J=6.9 Hz,3H), 1.14 (d, J=6.9 Hz, 3H).

The mixture of6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileatropisomers (10.59 g, “Intermediate B”, several batches) was purifiedby Chiral-Prep-HPLC with the following conditions: Column, CHIRALPAK IC,3.0×100 mm, 3 μm; mobile phase, IPA/ACN=1/1(V/V); Detection wavelength,UV 210 nm. This resulted in 4.99 g (47%) of6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile (the first eluting isomer, “Intermediate B-1”, M or Patropisomer) as a brown solid;

¹H NMR (400 MHz, CD₃OD) δ 8.48 (s, 1H), 8.44 (d, J=5.0 Hz, 1H),7.33-7.22 (m, 1H), 2.76-2.61 (m, 1H), 2.03 (s, 3H), 1.19 (d, J=6.8 Hz,3H), 1.06 (d, J=6.8 Hz, 3H).

And 4.60 g (43%) of6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the second eluting isomer, “Intermediate B-2”, P or M atropisomer) as abrown solid;

¹H NMR (400 MHz, CD₃OD) δ 8.49 (s, 1H), 8.44 (d, J=5.0 Hz, 1H), 7.28 (d,J=5.0 Hz, 1H), 2.76-2.63 (m, 1H), 2.03 (s, 3H), 1.19 (d, J=6.8 Hz, 3H),1.079-1.00 (m, 3H).

Step 2. tert-butyl(R)-4-(6,7-dichloro-3-cyano-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-2-methylpiperazine-1-carboxylate(single isomer)

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile (the first eluting isomer in step 1) (1.05 g, 2.71 mmol),POCl₃ (2.04 g, 13.27 mmol), DIEA (3.09 g, 23.93 mmol) and acetonitrile(20 mL). The mixture was stirred at 80° C. for 2 h. The reaction wascooled to room temperature and concentrated under vacuum. This resultedin4,6,7-trichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrilewhich was used directly in the next step.

Into a 100-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed4,6,7-trichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(crude) and acetonitrile (10 mL), DIEA (3.09 g, 23.93 mmol) andtert-butyl (R)-2-methylpiperazine-1-carboxylate (507 mg, 2.53 mmol) wereadded. The reaction mixture was stirred for 2 h at room temperature. Thereaction was then quenched by the addition of water (50 mL). Theresulting solution was extracted with ethyl acetate (3×50 mL), theorganic layers were combined and washed with brine (50 mL), dried overanhydrous Na₂SO₄, filtered and concentrated under vacuum. The residuewas purified by silica gel column eluted with EA/hexane (v/v=5/4). Thisresulted in 0.95 g (65% in two steps) of tert-butyl(R)-4-(6,7-dichloro-3-cyano-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-2-methylpiperazine-1-carboxylate(single isomer, P or M) as red solid. LCMS: m/z=571 [M+1]⁺.

Step 3.(R)-4-(4-acryloyl-3-methylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed tert-butyl(R)-4-(6,7-dichloro-3-cyano-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)-2-methylpiperazine-1-carboxylate(P or M isomer, 1.01 g, 1.767 mmol), DCM (8 mL) and TFA (4.605 g, 40.386mmol). The reaction mixture was stirred at room temperature for 1 h. Thereaction mixture was concentrated under vacuum. The residue wasdissolved by DCM (10 mL) into a 50 mL round-bottom flask, triethylamine(1.30 g, 12.85 mmol) was added. The reaction mixture was cooled to 0° C.and acryloyl chloride (0.23 g, 2.541 mmol) was added. The mixturestirred at room temperature for 0.5 h. The reaction was then quenched bythe addition of water (20 mL). The resulting solution was extracted withethyl acetate (3×20 mL), the organic layers were combined and washedwith brine (20 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under vacuum. The residues was purified by Prep-HPLCCH₃CN/H₂O (v/v=3/2). This resulted in 1.46 g (crude) of(R)-4-(4-acryloyl-3-methylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(P or M isomer) as yellow solid. LCMS: m/z=525 [M+1]⁺.

Step 4.4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(P or M isomer, “Compound 9”)

Into a 100 mL round bottom flask purged and maintained with an inertatmosphere of nitrogen was placed(R)-4-(4-acryloyl-3-methylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(P or M isomer, 1.64 g, 3.11 mmol),(2-amino-3,4,5,6-tetrafluorophenyl)boronic acid (1.37 g, 6.57 mmol),Pd(PPh₃)₄ (0.37 g, 315.86 mol), Na₂CO₃ (1.11 g, 10.49 mmol), 1,4-Dioxane(15 mL) and Water (2 mL). The resulting mixture was stirred at 80° C.for 30 min. Three batches of (2-amino-3,4,5,6-tetrafluorophenyl)boronicacid (2.84 g, 13.59 mmol) was added in 3 h. After the reaction wascompleted, the reaction was evaporated under vacuum and applied onto asilica gel column eluted with EA/Hexane (v/v=7/3), the crude waspurified by HPLC eluted with CH₃CN/H₂O (v/v=3/2), this resulted in (784mg, 38.50%) of7-(2-amino-3,4,5,6-tetrafluoro-phenyl)-6-chloro-1-(2-isopropyl-4-methyl-3-pyridyl)-4-[(3R)-3-methyl-4-prop-2-enoyl-piperazin-1-yl]-2-oxo-1,8-naphthyridine-3-carbonitrile(P or M isomer, “Compound 9”) as light yellow solid. LCMS: m/z=654[M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.48 (s, 1H), 8.42-8.31 (m, 1H), 7.23 (d,J=5.0 Hz, 1H), 6.76 (dd, J=16.7, 10.7 Hz, 1H), 6.20 (dd, J=16.7, 1.8 Hz,1H), 5.73 (dd, J=10.6, 1.9 Hz, 1H), 4.40 (s, 1H), 4.18-4.10 (m, 2H),3.94-3.83 (m, 2H), 3.65-3.54 (m, 2H), 2.64-2.51 (m, 1H), 2.06-1.92 (m,3H), 1.44-1.30 (m, 3H), 1.10-1.03 (m, 3H), 0.96-0.81 (m, 3H).

Example 104-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-6-chloro-3,4,5-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 10”)(P)-4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-6-chloro-3,4,5-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile;and(M)-4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-6-chloro-3,4,5-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Step 1.(R)-4-(4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-3,4,5-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Into a 20-mL round-bottom flask was placed(R)-4-(4-acryloyl-3-methylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(1.114 g, 2.12 mmol), (2-amino-3,4,5-trifluorophenyl)boronic acid (0.787g, 4.12 mmol), Pd(PPh₃)₄ (0.527 g, 0.45 mmol), Na₂CO₃ (0.652 g, 6.15mmol), dioxane (15 mL) and water (3 mL). The reaction mixture wasstirred at 80° C. for 1 h. The reaction mixture was filtered andconcentrated under vacuum. The residue was purified by Prep-HPLCCH₃CN/H₂O (0.05% NH₄HCO₃) (v/v=2/1). This resulted in 1.941 g (69%) of(R)-4-(4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-3,4,5-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileas yellow solid. LCMS: m/z=636 [M+1]⁺.

Step 2.4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-6-chloro-3,4,5-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 10”)

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed(R)-4-(4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-3,4,5-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(1.904 g, 3.00 mmol), NCS (0.946 g, 6.17 mmol) and AcOH (5 mL). Themixture was stirred at r.t. for 1 d. The residue was purified byPrep-HPLC (CH₃CN/H₂O)(v/v=6/4). This resulted in 0.497 g (25%) of4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-6-chloro-3,4,5-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 10”) as yellow solid. LCMS: m/z=670 [M+1]⁺.

Step 3.4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-6-chloro-3,4,5-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(first eluting isomer, “Compound 10-1”) &4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-6-chloro-3,4,5-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(second eluting isomer, “Compound 10-2”)

The mixture of4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-6-chloro-3,4,5-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileatropisomers (495 mg) was purified by Chiral-Prep-HPLC with thefollowing conditions: Column, CHIRAL ART Cellulose-SB, 3 cm×25 cm, 5 um;mobile phase, (Hex:DCM=3:1): EtOH (v/v=90:10); Detection wavelength, UV220 nm. This resulted in 204 mg (41.21%) of4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-6-chloro-3,4,5-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the first eluting isomer, “Compound 10-1”) as a yellow solid. LCMS:m/z=670 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.58 (s, 1H), 8.41 (dd, J=5.0, 1.4 Hz, 1H),7.23 (dd, J=8.4, 3.2 Hz, 1H), 6.85 (dd, J=16.7, 10.7 Hz, 1H), 6.29 (dd,J=16.8, 1.8 Hz, 1H), 5.82 (dd, J=10.6, 1.9 Hz, 1H), 4.88 (s, 2H), 4.60(s, 1H), 4.29-4.22 (m, 1H), 4.09-3.90 (m, 2H), 3.62 (s, 1H), 2.89-2.70(m, 1H), 1.95 (d, J=12.6 Hz, 3H), 1.45 (d, J=6.5 Hz, 3H), 1.18 (t, J=8.0Hz, 3H), 1.00 (dd, J=12.5, 6.8 Hz, 3H).

And 170 mg (28.34%) of4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-6-chloro-3,4,5-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the second eluting isomer, “Compound 10-2”) as a yellow solid. LCMS:m/z=670 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.59 (s, 1H), 8.41 (d, J=5.0 Hz, 1H),7.34-7.17 (m, 1H), 6.85 (dd, J=16.7, 10.7 Hz, 1H), 6.29 (dd, J=16.8, 1.8Hz, 1H), 5.82 (dd, J=10.6, 1.9 Hz, 1H), 4.88 (s, 2H), 4.60 (s, 1H),4.27-4.18 (m, 1H), 4.00 (s, 2H), 3.73-3.56 (m, 1H), 2.77-2.49 (m, 1H),2.05 (d, J=10.4 Hz, 3H), 1.46 (d, J=6.5 Hz, 3H), 1.16 (dd, J=6.8, 1.3Hz, 3H), 1.06-0.84 (m, 3H).

Example 114-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 11”)(P)-4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile;and(M)-4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Step 1.4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 11”)

Into a 20-mL round-bottom flask was placed(S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(0.311 g, 0.59 mmol), (2-amino-3,4,5,6-tetrafluorophenyl)boronic acid(0.512 g, 1.49 mmol), Pd(PPh₃)₄ (0.085 g, 0.073 mmol), Na₂CO₃ (0.126 g,1.19 mmol), dioxane (8 mL) and water (2 mL). The reaction mixture wasstirred at 80° C. for 1 h. The reaction mixture was filtered andconcentrated under vacuum. The residue was purified by Prep-HPLCCH₃CN/H₂O (0.05% NH₄HCO₃)(v/v=2/1). This resulted in 135 mg (35%) of4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileas yellow solid (“Compound 11”). LCMS: m/z=654 [M+1]⁺.

Step 2.4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(first eluting isomer, “Compound 11-1”) &4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(second eluting isomer, “Compound 11-2”)

The mixture of4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileatropisomers (135 mg) was purified by Chiral-Prep-HPLC with thefollowing conditions: Column, CHIRALPAK IF, 2 cm×25 cm, 5 um mobilephase, ((Hex:DCM=3:1):IPA=80:20; Detection wavelength, UV 220 nm. Thisresulted in 67 mg (49%) of4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the first eluting isomer, “Compound 11-1”) as a yellow solid. LCMS:m/z=654 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.52 (s, 1H), 8.43-8.44 (m, 1H), 7.26 (t,J=5.5 Hz, 1H), 7.02-6.77 (m, 1H), 6.33 (d, J=16.2 Hz, 1H), 5.85 (d,J=10.5 Hz, 1H), 4.67-4.39 (m, 2H), 4.17-4.18 (m, 3H), 3.78-3.56 (m, 2H),2.81-2.83 (m, 1H), 1.93-1.94 (m, 3H), 1.38-1.40 (m, 3H), 1.18-1.19 (m,3H), 1.01-1.04 (m, 3H).

And 61 mg (45%) of4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the second eluting isomer, “Compound 11-2”) as a yellow solid. LCMS:m/z=654 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.53 (s, 1H), 8.43-8.44 (m, 1H), 7.29-7.25 (m,1H), 6.85-6.86 (m, 1H), 6.33 (d, J=15.3 Hz, 1H), 5.85 (d, J=10.7 Hz,1H), 4.53-4.54 (m, 2H), 4.16-4.17 (m, 3H), 3.65-3.67 (m, 2H), 2.58-2.59(m, 1H), 2.16-2.02 (m, 3H), 1.38-1.40 (m, 3H), 1.19-1.11 (m, 3H),0.98-0.99 (m, 3H).

Example 124-(4-acryloylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 12”)(P)-4-(4-acryloylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile;and(M)-4-(4-acryloylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Step 1. 2,3,4-trifluoro-1-iodo-5-nitrobenzene

Into a 250-mL three-neck bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed 1,2,3-trifluoro-4-nitrobenzene(4.98 g, 28.12 mmol), N-iodosuccinimide (15.99 g, 71.07 mmol),trifluoromethanesulfonic acid (25 mL) and stirred. The resultingsolution was stirred overnight at room temperature. The reaction wasthen quenched by the addition of water (200 mL) slowly. The resultingsolution was extracted with EA (2×100 mL), the organic layers werecombined, washed with water (100 mL) and brine (100 mL), dried overanhydrous Na₂SO₄, the residue was concentrated under vacuum. Theresulting crude product was further purified by C₁₈ column eluted withACN/H₂O (v/v=0%˜100%). This resulted in 4.66 g (54.69% yield) of2,3,4-trifluoro-1-iodo-5-nitrobenzene.

Step 2. 2,3,4-trifluoro-5-iodoaniline

Into a 250-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed 2,3,4-trifluoro-1-iodo-5-nitrobenzene(4.66 g, 15.38 mmol), Fe iron (5.02 g, 89.93 mmol), ammonium chloride(8.57 g, 160.25 mmol), EtOH (30 mL) and water (30 mL). The reactionmixture was stirred at 55° C. for 1 h. The reaction was filtered and thefilter cake was washed with EA (2×20 mL). The organic layers werecombined and washed with brine (100 mL), dried over anhydrous Na₂SO₄.The resulting solution was concentrated under vacuum and applied onto asilica gel column eluted with EA/hexane (v/v=1/1). This resulted in 3.69g (87% yield) of 2,3,4-trifluoro-5-iodoaniline as yellow oil. LCMS:m/z=274 [M+1]⁺.

Step 3. (5-amino-2,3,4-trifluorophenyl)boronic acid

Into a 250-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed 2,3,4-trifluoro-5-iodoaniline (2.09 g, 7.67mmol), Bis(pinacolato)diboron (9.46 g, 37.25 mmol), Pd(dppf)Cl₂ (1.06 g,1.45 mmol), KOAc (5.10 g, 52.00 mmol) and NMP (80 mL). The reactionmixture was stirred at 95° C. for 2 h. The reaction was then quenched bythe addition of saturated sodium carbonate aqueous solution (100 mL).The reaction mixture was stirred at room temperature for 2 h. Theresulting solution was extracted with EA (3×100 mL), the organic layerswere combined, then washed with brine (100 mL), dried over anhydrousNa₂SO₄, the residue was concentrated under vacuum. The resulting crudewas further purified by C₁₈ column eluted with ACN/H₂O (v/v=30%˜50%).This resulted in 1.48 g (106% yield) of(5-amino-2,3,4-trifluorophenyl)boronic acid as yellow oil. LCMS: m/z=192[M+1]⁺.

Step 4.4-(4-acryloylpiperazin-1-yl)-7-(5-amino-2,3,4-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Into a 40-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed4-(4-acryloylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(0.68 g, 1.34 mmol), (5-amino-2,3,4-trifluorophenyl)boronic acid (0.51g, 2.67 mmol), Pd(PPh₃)₄ (0.31 g, 0.27 mmol), Na₂CO₃ (0.41 g, 3.86mmol), dioxane (20 mL) and water (4 mL). The reaction mixture wasstirred at 80° C. for 2 h. The reaction was then quenched by theaddition of water (50 mL) and ethyl acetate (20 mL). The resultingsolution was extracted with ethyl acetate (1×50 mL), the organic layerswere combined and dried over anhydrous Na₂SO₄, filtered and concentratedunder vacuum. The resulting crude product was further purified by C₁₈column eluted with ACN/H₂O (v/v=0%˜60%). This resulted in 0.15 g (18%yield) of4-(4-acryloylpiperazin-1-yl)-7-(5-amino-2,3,4-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileas yellow solid. LCMS: m/z=622 [M+1]⁺.

Step 5.4-(4-acryloylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 12”)

Into a 8-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed4-(4-acryloylpiperazin-1-yl)-7-(5-amino-2,3,4-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(0.15 g, 0.25 mmol), NCS (0.10 g, 0.77 mmol) and AcOH (4 mL). Themixture was stirred for overnight at room temperature. The reaction wasthen quenched by the addition of water (50 mL). The resulting solutionwas extracted with ethyl acetate (2×50 mL), the organic layers werecombined and washed with saturated sodium bicarbonate aqueous solution(2×50 mL), dried over anhydrous Na₂SO₄, filtered and concentrated undervacuum. The resulting crude product was further purified by C₁₈ columneluted with ACN/H₂O (0.15% NH₄HCO₃) (v/v=30%˜80%). This resulted in 21mg (13% yield) of4-(4-acryloylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 12”) as yellow solid. LCMS: m/z=656 [M+1]⁺.

¹H NMR (400 MHz, DMSO-d₆) δ 8.57 (s, 1H), 8.44 (d, J=4.7 Hz, 1H), 7.23(t, J=4.6 Hz, 1H), 6.92 (dd, J=16.7, 10.5 Hz, 1H), 6.20 (dd, J=16.6, 2.3Hz, 1H), 5.86 (s, 2H), 5.77 (dd, J=10.4, 2.2 Hz, 1H), 4.29-3.50 (m, 8H),3.72-3.66 (m, 1H), 1.89 (d, J=10.7 Hz, 3H), 1.08 (dd, J=6.6, 2.4 Hz,3H), 0.97-0.77 (m, 3H).

Step 6.4-(4-acryloylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(first eluting isomer, “Compound 12-1”) &4-(4-acryloylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile (second eluting isomer, “Compound 12-2”)

The mixture of4-(4-acryloylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileatropisomers (110 mg, several batches) was purified by Chiral-Prep-HPLCwith the following conditions: Column, CHIRAL ART Cellulose-SB, 3 cm×25cm, 5 um; mobile phase, (Hex:DCM=3:1): EtOH (v/v=95:5); Detectionwavelength, UV 220 nm. This resulted in 53 mg (48.18%) of4-(4-acryloylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the first eluting isomer, “Compound 12-1”) as a yellow solid. LCMS:m/z=656 [M+1]⁺.

¹H NMR (400 MHz, DMSO-d₆) δ 8.57 (s, 1H), 8.44 (dd, J=4.9, 1.5 Hz, 1H),7.24 (t, J=4.8 Hz, 1H), 6.92 (dd, J=16.7, 10.4 Hz, 1H), 6.21 (dd,J=16.6, 2.4 Hz, 1H), 5.88 (s, 2H), 5.77 (dd, J=10.4, 2.4 Hz, 1H),4.25-2.64 (d, J=27.7 Hz, 8H), 2.78-2.60 (m, 1H), 1.89 (d, J=10.4 Hz,3H), 1.12-1.01 (m, 3H), 0.97-0.83 (m, 3H).

And 51 mg (46.36%) of4-(4-acryloylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the second eluting isomer, “Compound 12-2”) as a yellow solid. LCMS:m/z=656 [M+1]⁺.

¹H NMR (400 MHz, DMSO-d₆) δ 8.57 (s, 1H), 8.44 (d, J=4.9 Hz, 1H), 7.24(t, J=4.7 Hz, 1H), 6.92 (dd, J=16.6, 10.4 Hz, 1H), 6.21 (dd, J=16.6, 2.5Hz, 1H), 5.88 (s, 2H), 5.77 (dd, J=10.3, 2.4 Hz, 1H), 3.89 (d, J=25.8Hz, 8H), 2.73-2.64 (m, 1H), 1.89 (d, J=10.5 Hz, 3H), 1.08 (dd, J=6.8,2.6 Hz, 3H), 0.90 (dd, J=19.2, 6.7 Hz, 3H).

Example 134-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 13”)(P)-4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile;and(M)-4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Step 1.4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 13”)

Into a 20-mL round-bottom flask was placed4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(0.312 g, 0.58 mmol),2,3,4,5-tetrafluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline(0.601 g, 2.07 mmol), Pd(pph₃)₄ (0.125 g, 0.11 mmol), Na₂CO₃ (0.189 g,1.78 mmol), dioxane (8 mL) and water (2 mL). The reaction mixture wasstirred at 80° C. for 1 h. The reaction mixture was filtered andconcentrated under vacuum. The residue was purified by Prep-HPLCCH₃CN/H₂O (0.05% NH₄HCO₃)(v/v=2/1). This resulted in 0.132 g (34%) of4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 13”) as yellow solid. LCMS: m/z=668 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.85 (s, 1H), 8.67 (d, J=5.8 Hz, 1H), 7.85 (d,J=5.9 Hz, 1H), 6.94-6.81 (m, 1H), 6.33 (d, J=16.6 Hz, 1H), 5.84 (d,J=10.8 Hz, 1H), 4.78 (s, 2H), 4.09-3.95 (m, 2H), 3.95-3.81 (m, 2H),3.13-2.99 (m, 1H), 2.33-2.18 (m, 3H), 1.68-1.59 (m, 6H), 1.36-1.24 (m,3H), 1.23-1.09 (m, 3H).

Step 2.4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(first eluting isomer, “Compound 13-1”) &4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(second eluting isomer, “Compound 13-2”)

The mixture of4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileatropisomers (930 mg, several batches) was purified by Chiral-Prep-HPLCwith the following conditions: Column, CHIRALPAK IA, 3 cm×25 cm, 5 um;mobile phase, Hex:EtOH=80:20; Detection wavelength, UV 220 nm. Thisresulted in 424 mg (45.59%) of4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the first eluting isomer, “Compound 13-1”) as a yellow solid. LCMS:m/z=668 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.83 (s, 1H), 8.52 (d, J=5.3 Hz, 1H), 7.47 (d,J=5.2 Hz, 1H), 6.88 (dd, J=16.7, 10.6 Hz, 1H), 6.33 (dd, J=16.7, 1.9 Hz,1H), 5.84 (dd, J=10.6, 1.9 Hz, 1H), 4.77 (s, 2H), 4.10-3.94 (m, 2H),3.91-3.79 (m, 2H), 2.81-2.83 (m, 1H), 2.08-2.10 (m, 3H), 1.65 (t, J=6.0Hz, 6H), 1.26-1.19 (m, 3H), 1.01-1.02 (m, 3H).

And 372 mg (40.00%) of4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the second eluting isomer, “Compound 13-2”) as a yellow solid. LCMS:m/z=668 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.83 (s, 1H), 8.52 (d, J=5.3 Hz, 1H), 7.47 (d,J=5.2 Hz, 1H), 6.88 (dd, J=16.7, 10.6 Hz, 1H), 6.33 (dd, J=16.7, 1.9 Hz,1H), 5.84 (dd, J=10.6, 1.9 Hz, 1H), 4.77 (s, 2H), 4.10-3.94 (m, 2H),3.91-3.79 (m, 2H), 2.82-2.83 (m, 1H), 2.09-2.11 (m, 3H), 1.65 (t, J=6.0Hz, 6H), 1.26-1.19 (m, 3H), 1.04-1.06 (m, 3H).

Example 144-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 14”)

Step 1.4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 14”)

Into a 20-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed(R)-4-(4-acryloyl-3-methylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(353 mg, 0.67 mmol),2,3,4,5-tetrafluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline(604 mg, 2.08 mmol), Pd(PPh₃)₄ (185 mg, 0.16 mmol), Na₂CO₃ (235 mg, 2.22mmol), dioxane (7 mL) and water (0.7 mL). The reaction mixture wasstirred at 80° C. for 1 h. The reaction mixture was filtered andconcentrated under vacuum. The reaction mixture was concentrated undervacuum. The residue was purified by Prep-HPLC (CH₃CN/H₂O (v/v=7/3)).This resulted in 29 mg (7% yield) of4-((R)-4-acryloyl-3-methylpiperazin-1-yl)-7-(2-amino-3,4,5,6-tetrafluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 14”) as yellow solid. LCMS: m/z=654 [M+1]⁺. 5 ¹H NMR (400MHz, CD₃OD) δ 8.50 (d, J=52.4 Hz, 2H), 7.27 (s, 1H), 6.96-6.66 (m, 1H),6.29 (d, J=16.2 Hz, 1H), 5.83 (d, J=9.6 Hz, 1H), 4.70-4.43 (m, 1H),4.18-4.08 (m, 4H), 3.70-3.60 (m, 2H), 2.82-2.48 (m, 1H), 2.03 (s, 3H),1.48-1.40 (m, 3H), 1.24-0.84 (m, 6H).

Example 154-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 15”)(P)-4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile;and(M)-4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Step 1.(S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-7-(5-amino-2,3,4-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Into a 20-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen was placed(S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(0.310 g, 0.59 mmol), (5-amino-2,3,4-trifluorophenyl)boronic acid (0.285g, 1.49 mmol), Pd(PPh₃)₄ (0.085 g, 0.073 mmol), Na₂CO₃ (0.126 g, 1.19mmol), dioxane (8 mL) and water (2 mL). The reaction mixture was stirredat 80° C. for 1 h. The reaction mixture was filtered and concentratedunder vacuum. The residue was purified by Prep-HPLC CH₃CN/H₂O (0.05%NH₄HCO₃)(v/v=2/1). This resulted in 135 mg (35%) of(S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-7-(5-amino-2,3,4-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile asyellow solid. LCMS: m/z=636 [M+1]⁺.

Step 2.4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 15”)

Into a 8-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed(S)-4-(4-acryloyl-2-methylpiperazin-1-yl)-7-(5-amino-2,3,4-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(0.123 g, 193.376 mol), NCS (0.053 g, 396.906 mol) and AcOH (2 mL). Themixture was stirred for overnight at room temperature.

The reaction was then quenched by the addition of saturated sodiumbicarbonate aqueous solution (100 mL). The resulting solution wasextracted with ethyl acetate (3×100 mL), the organic layers werecombined and concentrated under vacuum. The resulting crude product wasfurther purified by C₁₈ column eluted with ACN/H₂O (0.15% NH₄HCO₃)(v/v=30%˜80%). This resulted in 55 mg (42% yield) of4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound 15”) as yellow solid. LCMS: m/z=670 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.54 (s, 1H), 8.41 (d, J=4.8 Hz, 1H), 7.24(dd, J=11.5, 5.9 Hz, 1H), 6.98-6.74 (m, 1H), 6.33 (d, J=16.3 Hz, 1H),5.84 (d, J=10.5 Hz, 1H), 4.69-3.97 (m, 5H), 3.78-3.52 (m, 2H), 2.86-2.56(m, 1H), 2.12-1.88 (m, 3H), 1.47-1.30 (m, 3H), 1.25-1.10 (m, 3H),1.07-0.88 (m, 3H).

Step 2.4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(first eluting isomer, “Compound 15-1”) &4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(second eluting isomer, “Compound 15-2”)

The mixture of4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileatropisomers (53 mg) was purified by Chiral-Prep-HPLC with the followingconditions: Column, CHIRALPAK IF, 2 cm×25 cm, 5 um mobile phase,(Hex:DCM=3:1):EtOH (v/v=9:1); Detection wavelength, UV 220 nm. Thisresulted in 20 mg (49%) of4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the first eluting isomer. “Compound 15-1”) as a yellow solid. LCMS:m/z=670 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.56 (s, 1H), 8.43 (d, J=5.0 Hz, 1H), 7.28 (t,J=4.9 Hz, 1H), 6.99-6.79 (m, 1H), 6.35 (d, J=18.0 Hz, 1H), 5.87 (d,J=10.7 Hz, 1H), 4.68-4.06 (m, 5H), 3.78-3.56 (m, 2H), 2.69-2.56 (m, 1H),2.09 (d, J=9.8 Hz, 3H), 1.42 (s, 3H), 1.17 (d, J=6.8 Hz, 3H), 0.98 (dd,J=18.6, 6.8 Hz, 3H).

And 17 mg (45%) of4-((S)-4-acryloyl-2-methylpiperazin-1-yl)-7-(3-amino-2-chloro-4,5,6-trifluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the second eluting isomer, “Compound 15-2”) as a yellow solid. LCMS:m/z=670 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.56 (s, 1H), 8.44 (d, J=5.0 Hz, 1H), 7.26 (t,J=5.4 Hz, 1H), 6.87 (d, J=27.5 Hz, 1H), 6.97-6.80 (m, 1H), 5.87 (d,J=11.5 Hz, 1H), 4.65-4.05 (m, 5H), 3.78-3.57 (m, 2H), 2.88-2.78 (m, 1H),1.95 (d, J=12.5 Hz, 3H), 1.44-1.34 (m, 3H), 1.22 (d, J=6.7 Hz, 3H), 1.03(dd, J=11.9, 6.8 Hz, 3H).

The following compounds can be synthesized following the similar methodsas described in the above-mentioned examples:

Example A4-(4-acryloylpiperazin-1-yl)-7-(2-amino-6-fluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound A”)

Step 1.4,6,7-trichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed6,7-dichloro-4-hydroxy-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(980 mg, 2.51 mmol), POCl₃ (1150 mg, 7.50 mmol), DIEA (1.32 g, 10.21mmol) and acetonitrile (12 mL). The mixture was stirred at 80° C. for 2h. The reaction was cooled to room temperature and concentrated undervacuum. This resulted in4,6,7-trichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrilewhich was used directly in the next step.

Step 2. tert-butyl4-(6,7-dichloro-3-cyano-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazine-1-carboxylate

Into a 100-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed4,6,7-trichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(1.20 g, crude), acetonitrile (20 mL), DIEA (660 mg, 5.10 mmol) andtert-butyl piperazine-1-carboxylate (0.57 g, 3.06 mmol). The reactionmixture was stirred for 2 h at room temperature. The reaction was thenquenched by the addition of water (50 mL). The resulting solution wasextracted with ethyl acetate (3×50 mL), the organic layers were combinedand washed with brine (50 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under vacuum. The residues was purified by silica gelcolumn eluted with EA/hexane (v/v=30%˜70%). This resulted in 0.92 g (65%in two steps) of tert-butyl4-(6,7-dichloro-3-cyano-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazine-1-carboxylateas yellow solid. LCMS: m/z=557 [M+1]⁺.

Step 3.4-(4-acryloylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Into a 50-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed tert-butyl4-(6,7-dichloro-3-cyano-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridin-4-yl)piperazine-1-carboxylate(920 mg, 1.65 mmol), TFA (4 ml) and DCM (15 mL). The reaction mixturewas stirred at room temperature for 2 h. The reaction mixture wasconcentrated under vacuum. The residue was dissolved by DCM (15 mL) in50-mL round-bottom flask, which was followed by the added of DIEA (1.02g, 10.08 mmol). The reaction mixture was cooled to 0° C. and acryloylchloride (190 mg, 2.09 mmol) was added. The mixture stirred at roomtemperature for 2 h. The reaction was then quenched by the addition ofwater (30 mL). The resulting solution was extracted with ethyl acetate(3×50 mL), the organic layers were combined and washed with brine (30mL), dried over anhydrous Na₂SO₄, filtered and concentrated undervacuum. The residue was purified by silica gel column eluted withEA/hexane (V/V=40%˜80%). This resulted in 0.86 g (crude) of4-(4-acryloylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileas yellow solid. LCMS: m/z=511 [M+1]⁺.

Step 4.4-(4-acryloylpiperazin-1-yl)-7-(2-amino-6-fluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound A”)

Into a 20-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed4-(4-acryloylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(99 mg, 0.19 mmol),3-fluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (78 mg,0.33 mmol), Pd(PPh₃)₄ (44 mg, 0.03 mmol), Na₂CO₃ (65 mg, 0.61 mmol),dioxane (4 mL) and water (1 mL). The reaction mixture was stirred at 90°C. for 2 h. The reaction mixture was filtered and concentrated undervacuum. The residue was purified by Prep-HPLC CH₃CN/H₂O (v:v=1/1)). Thisresulted in 12 mg (10%) of4-(4-acryloylpiperazin-1-yl)-7-(2-amino-6-fluorophenyl)-6-chloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Compound A”) as yellow solid. LCMS: m/z=586 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.52 (s, 1H), 8.46 (d, J=5.0 Hz, 1H), 7.29 (d,J=4.7 Hz, 1H), 7.11 (d, J=6.6 Hz, 1H), 6.89 (dd, J=16.7, 10.6 Hz, 1H),6.56-6.49 (m, 1H), 6.38-6.30 (m, 2H), 5.90-5.81 (m, 1H), 4.18-3.84 (m,8H), 2.82-2.70 (m, 1H), 2.08-1.98 (m, 3H), 1.19 (t, J=7.1 Hz, 3H),1.10-0.93 (m, 3H).

Amgen 64-(4-acryloylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Amgen 6”)

Step 1.4-(4-acryloylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Into a 8-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed4-(4-acryloylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(75 mg, 0.15 mmol), (2-fluorophenyl)boronic acid (56 mg, 0.40 mmol),Pd(dppf)Cl₂ (25 mg, 34.17 μmol), Na₂CO₃ (69 mg, 0.65 mmol), dioxane (1mL) and water (0.2 mL). The reaction mixture was stirred at 90° C. for 2h. The reaction mixture was filtered and concentrated under vacuum. Theresidue was purified by silica gel column eluted with EA/hexane(v/v=7/3). The collecting fluid concentrated under vacuum. The residuewas purified by Prep-HPLC CH₃CN/H₂O (v/v=3/2). This resulted in 22 mg of4-(4-acryloylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Amgen 6”) as yellow solid. LCMS: m/z=571 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.52 (s, 1H), 8.43 (d, J=5.0 Hz, 1H),7.54-7.46 (m, 1H), 7.32-7.13 (m, 4H), 6.89 (dd, J=16.7, 10.6 Hz, 1H),6.33 (d, J=16.7 Hz, 1H), 5.86 (d, J=10.6 Hz, 1H), 4.15-3.90 (m, 8H),2.79-2.65 (m, 1H), 2.04 (s, 3H), 1.19 (d, J=6.8 Hz, 3H), 1.00 (d, J=6.7Hz, 3H).

The mixture of4-(4-acryloylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileatropisomers (1.59 g) was purified by Chiral-Prep-HPLC with thefollowing conditions: Column, CHIRAL Cellulose-SB, 3 cm×25 cm, 5 um;mobile phase, CO₂,IPA:ACN=1:1; Detector, UV 254 nm. This resulted in 739mg (46%) of4-(4-acryloylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the first eluting isomer, “Amgen 6-1”) as a yellow solid; LCMS: m/z=571[M+1]⁺.

And 709 mg (45%) of4-(4-acryloylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile (thesecond eluting isomer, “Amgen 6-2”) as a yellow solid; LCMS: m/z=571[M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.50 (s, 1H), 8.41 (d, J=5.0 Hz, 1H),7.52-7.37 (m, 1H), 7.30-7.08 (m, 4H), 6.87 (dd, J=16.8, 10.6 Hz, 1H),6.38-6.24 (m, 1H), 5.83 (dd, J=10.6, 1.8 Hz, 1H), 4.09-3.82 (m, 8H),2.78-2.63 (m, 1H), 2.02 (s, 3H), 1.16 (d, J=6.8 Hz, 3H), 0.98 (d, J=6.8Hz, 3H).

Amgen 6.34-(4-acryloylpiperazin-1-yl)-7-(2-amino-6-fluorophenyl)-6-chloro-1-(4,6-diisopropylpyrimidine-5-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Amgen 6.3”)

4-(4-acryloylpiperazin-1-yl)-7-(2-amino-6-fluorophenyl)-6-chloro-1-(4,6-diisopropylpyrimidin-5-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile (“Amgen 6.3”) wereprepared according to prior method as yellow solid. LCMS: m/z=615[M+1]⁺.

¹H NMR (400 MHz, DMSO-d₆) δ 9.09 (s, 1H), 8.48 (s, 1H), 7.05 (q, J=7.8Hz, 1H), 6.93 (dd, J=16.6, 10.4 Hz, 1H), 6.44 (d, J=8.3 Hz, 1H), 6.31(t, J=8.9 Hz, 1H), 6.21 (dd, J=16.7, 2.4 Hz, 1H), 5.77 (dd, J=10.4, 2.4Hz, 1H), 5.08 (s, 2H), 3.90 (m, 8H), 2.90-2.74 (m, 1H), 2.70-2.55 (m,1H), 1.07 (dd, J=12.2, 6.7 Hz, 6H), 1.00 (d, J=6.6 Hz, 3H), 0.86 (d,J=6.7 Hz, 3H).

Amgen 7.34-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Amgen 7.3”)

Step 1.4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile

Into a 20-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6,7-dichloro-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile (109 mg, 0.20 mmol),((2-fluorophenyl)boronic acid (110 mg, 0.79 mmol), Pd(PPh₃)₄ (85 mg,0.073 mmol), Na₂CO₃ (69 mg, 0.65 mmol), dioxane (6 mL) and water (2 mL).The reaction mixture was stirred at 80° C. for 4 h. The reaction wasthen quenched by the addition of water (100 mL). The resulting solutionwas extracted with ethyl acetate (3×100 mL), the organic layers werecombined and washed with brine (100 mL), dried over anhydrous Na₂SO₄,filtered and concentrated under vacuum. The residue was purified byPrep-HPLC (CH₃CN/H₂O (v/v=1/1). This resulted in 31 mg (26% in twosteps) of4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(“Amgen 7.3”) as yellow solid. LCMS: m/z=599 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.80 (s, 1H), 8.44 (d, J=5.0 Hz, 1H), 7.48(dd, J=8.4, 4.6 Hz, 1H), 7.40-7.22 (m, 4H), 6.98-6.85 (m, 1H), 6.35 (d,J=16.5 Hz, 1H), 5.86 (d, J=10.6 Hz, 1H), 4.79 (s, 2H), 4.14-3.79 (m,4H), 2.80-2.74 (m, 1H), 2.05 (s, 3H), 1.68 (d, J=7.0 Hz, 6H), 1.19 (d,J=6.8 Hz, 3H), 1.01 (d, J=6.7 Hz, 3H).

The mixture of4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrileatropisomers (1.48 g) was purified by Chiral-Prep-HPLC with thefollowing conditions: Column, CHIRAL Cellulose-SB, 3 cm×25 cm, 5 um;mobile phase, Hex/EtOH=(v/v=50/50); Detection wavelength, UV 254 nm.This resulted in 625 mg (42%) of4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the first eluting isomer, “Amgen 7.3-1”) as a yellow solid; LCMS:m/z=599 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.68 (s, 1H), 8.32 (d, J=5.0 Hz, 1H),7.42-7.30 (m, 1H), 7.23-7.01 (m, 4H), 6.78 (dd, J=16.7, 10.6 Hz, 1H),6.22 (dd, J=16.7, 1.9 Hz, 1H), 5.73 (dd, J=10.6, 1.9 Hz, 1H), 4.66 (s,2H), 3.99-3.83 (m, 2H), 3.74-3.72 (m, 2H), 2.63-2.61 (m, 1H), 1.93 (s,3H), 1.55 (d, J=7.0 Hz, 6H), 1.10-1.08 (m, 3H), 0.90-0.89 (m, 3H).

And 669 mg (45%) of4-((3S,5R)-4-acryloyl-3,5-dimethylpiperazin-1-yl)-6-chloro-7-(2-fluorophenyl)-1-(2-isopropyl-4-methylpyridin-3-yl)-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carbonitrile(the second eluting isomer, “Amgen 7.3-2”) as a yellow solid; LCMS:m/z=599 [M+1]⁺.

¹H NMR (400 MHz, CD₃OD) δ 8.80 (s, 1H), 8.43 (d, J=5.0 Hz, 1H),7.57-7.41 (m, 1H), 7.37-7.07 (m, 4H), 6.90 (dd, J=16.7, 10.6 Hz, 1H),6.34 (dd, J=16.7, 1.9 Hz, 1H), 5.85 (dd, J=10.6, 1.9 Hz, 1H), 4.78 (s,2H), 4.07-3.98 (m, 2H), 3.86-3.84 (m, 2H), 2.76-2.74 (m, 1H), 2.05 (s,3H), 1.68-1.66 (m, 6H), 1.12-1.10 (m, 3H), 0.91-0.89 (m, 3H).

Pharmacological Testing 1. SOS1 Catalyzed Nucleotide Exchange Assay

HIS-KRAS (G12C, aa 2-185, Sino biological) was diluted to 5 μM in EDTAbuffer (20 mM HEPES, pH 7.4, 50 mM NaCl, 10 mM EDTA, 0.01% (v/v)Tween-20) and incubated for 30 min at 25° C. The EDTA pretreatedHIS-KRAS (G12C) was diluted to 12 nM in assay buffer (25 mM HEPES, pH7.4, 120 mM NaCl, 5 mM MgCl₂, 1 mM DTT, 0.01% (v/v) Tween 20, 0.1% (w/v)BSA) containing 120 nM GDP(Sigma) and MAb Anti 6HIS-Tb cryptateGold(Cisbio) and incubated for 1 hour at 25° C. to prepare GDP-loadedHIS-KRAS (G12C). The GDP-loaded HIS-KRAS (G12C) was pre-incubation withdiluted compounds in a 384-well plate (Greiner) for 1 hour, thenpurified SOS1 ExD (Flag tag, aa 564-1049) and BODIPY™ FL GTP(Invitrogen) were added to the assay wells (Final concentration: 3 nMHIS-KRAS (G12C), 2 μM SOS1 ExD, 80 nM BODIPY™ FL GTP, 21 ng/mL MAb Anti6HIS-Tb cryptate Gold) and incubated for 4 hours at 25° C. TR-FRETsignals were then read on Tecan Spark multimode microplate reader. Theparameters were F486: Excitation 340 nm, Emission 486 nm, Lag time 100s, Integration time 200 s; F515: Excitation 340 nm, Emission 515 nm, Lagtime 100 s, Integration time 200 s. TR-FRET ratios for each individualwells were calculated by equation: TR-FRET ratio=(Signal F515/SignalF486)*10000. Then the data were analyzed using a 4-parameter logisticmodel to calculate IC₅₀ values. The results of the SOS1 catalyzednucleotide exchange assay are in the following Table 8:

TABLE 8 SOS1 catalyzed nucleotide Compound exchange IC₅₀(nM) Compound 13.06 Compound 1-1 14.1 Compound 1-2 1.41 Compound 2 6.66 Compound 3 3.67Compound 4 3.48 Compound 5 5.97 Compound 6 1.51 Compound 7 14.9 Compound8 6.68 Compound 9 2.30 Compound 10-1 43.9 Compound 10-2 4.16 Compound11-1 29.6 Compound 11-2 2.11 Compound 12 3.85 Compound 12-1 16.0Compound 12-2 2.01 Compound 13 6.63 Compound 13-1 2.73 Compound 13-211.5 Compound 14 3.56 Compound 15-1 6.87 Compound 15-2 83.9 Compound A1.69 Amgen 6 4.40 Amgen 6.3 2.77 Amgen 7.3 10.5

From the Table 8, it can be seen that the representative compounds inthe present invention have better activity to inhibit the SOS1 catalyzednucleotide exchange.

2. Phospho-ERK1/2(THR202/TYR204) HTRF Assay

NCI-H358 cells expressing KRAS G12C were cultured in RPMI 1640 medium(Gibco) containing 10% fetal bovine serum (Gibco). The NCI-H358 cells inculture medium were seeded in 96-well plates at a concentration of40,000 cells/well and then put in a 37° C./5% CO₂ cell incubator toincubate overnight. The next day, culture medium was removed and thecompound diluted in assay medium (RPMI 1640, 0.1% FBS) was added in eachwell. After 2 hours incubation in a 37° C./5% CO₂ cell incubator, theassay medium in 96-well plates was removed, then 50 μL of 1× blockingreagent-supplemented lysis buffer (Cisbio) was added and the plates wereincubated at 25° C. for 45 min with shaking. 10 μL of cell lysates fromthe 96-well plates were transferred to a 384-well plate (Greiner)containing 2.5 μL/well HTRF® pre-mixed antibodies (Cisbio 64AERPEH).Incubate 4 hours at 25° C. and then read HTRF signals on Tecan Sparkmultimode microplate reader. The data were analyzed using a 4-parameterlogistic model to calculate IC₅₀ values. The results of thePhospho-ERK1/2(THR202/TYR204) HTRF assay are in the following Table 9:

TABLE 9 Compound p-ERK IC₅₀(nM) Compound 1 16.4 Compound 1-1 176Compound 1-2 10.2 Compound 2 33.6 Compound 3 21.4 Compound 4 40.1Compound 5 141 Compound 6 25.1 Compound 8 48.0 Compound 9 26.3 Compound10-1 217 Compound 10-2 22.0 Compound 11-1 258 Compound 11-2 23.6Compound 12 39.7 Compound 12-1 117 Compound 12-2 12.3 Compound 13 87.0Compound 13-1 21.8 Compound 13-2 138 Compound 14 37.6 Compound 15-1 44.0Compound 15-2 327 Compound A 14.2 Amgen 6 29.9 Amgen 6.3 20.1 Amgen 7.344.5

From Table 9, it can be seen that representative compounds in thepresent invention have better activity to inhibit the phosphorylation ofERK1/2 the NCI-H358 cells.

3. Mouse Pharmacokinetic Study

The purpose of this study was to evaluate the pharmacokinetic propertiesof compounds in Balb/c mouse (

) following single dose administration. The day before administration,mice were fasted overnight and free access to water. Six mice wereneeded for each compound and the six mice were divided into two groups(n=3/group), group A and group B. Mice in group A were treated with asingle 3 mg/kg dose of compound (iv). Mice in group B were treated witha single 10 mg/kg dose of compound (po). For each mouse in group A,blood samples were collected at pre-dose, and at the time point of0.083, 0.25, 0.5, 1, 2, 4, 6, 8 and 24 h post-dose. For each mouse ingroup B, blood samples were collected at pre-dose, and at the time pointof 0.25, 0.5, 1, 2, 3, 4, 6, 8 and 24 h post-dose. Blood samples wereplaced on ice until centrifugation to obtain plasma samples. The plasmasamples were stored at −80° C. until analysis. The concentration ofcompound in plasma samples were determined using a LC-MS/MS method. Theresults are in the following Table 10:

TABLE 10 3 mg/kg, iv 10 mg/kg, po CL (mL/ Vss C_(max) AUC_(0-24h) OralCompound min/kg) (L/kg) (ng/mL) (ng · h/mL) BA (F %) Compound 1 21.41.87 1397 4025 51.3 Compound 11 7.2 1.0 3340 9587 38 Compound 13 9.8 1.43013 12763 74 Compound 14 10 0.9 4023 10498 63 Compound A 65.2 1.13 241207 8.05 Amgen 6 72 2.6 901 1032 44 Amgen 6.3 32 1.4 1587 1658 32

From Table 10, it can be seen that Compound 1, Compound 11, Compound 13and Compound 14 have excellent pharmacokinetic properties (such as thehigher C_(max) and AUC) in mouse model comparative with the Compound A,Amgen 6 and Amgen 6.3, which make them more suitable for treatingcancers with KRAS G12C mutation as an orally therapeutic activeingredient in clinic.

4. Dog Pharmacokinetic Study

The purpose of this study was to evaluate the pharmacokinetic propertiesof compounds in beagle dog following single dose administration. The daybefore administration, dogs were fasted overnight and free access towater. Four beagle dogs were needed for each compound and the four dogswere divided into two groups (one male (

) and one female (

) in each group). Dogs in group A were treated with a single 1 mg/kgdose of compound (iv). Dogs in group B were treated with a single 10mg/kg dose of compound (po). For dogs in group A, blood samples werecollected at pre-dose, and at the time point of 0.083, 0.25, 0.5, 1, 2,4, 6, 8 and 24 h. For dogs in group B, blood samples were collected atpre-dose, and at the time point of 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hpost-dose. Blood samples were placed on ice until centrifugation toobtain plasma samples. The plasma samples were stored at −80° C. untilanalysis. The concentration of compound in plasma samples weredetermined using a LC-MS/MS method. The results are in following Table11:

TABLE 11 1 mg/kg, iv 10 mg/kg, po CL Vss C_(max) AUC_(0-24 h) Oral BACompound Sex (mL/min/kg) (L/kg) (ng/mL) (ng · h/mL) (F %) Compound 1-2

15.9 0.9 3420 6724 64.3

18.1 1.1 2980 5953 65.1 Compound 9

17.7 1.37 2490 7788 83.2

20.5 1.72 2260 6323 78.5 Amgen 6

11.8 0.412 3690 2944 20.9

15.5 0.543 2580 1326 12.4 Amgen 6.3

239 2.3 8.0 5.1 0.7

329 3.1 — — —

From Table 11, it can be seen that Compound 1-2 and Compound 9 haveexcellent pharmacokinetic properties (such as the higher C_(max) andAUC) in beagle dog model comparative with the Amgen 6 and 6.3, whichmake them more suitable for treating cancers with KRAS G12C mutation asan orally therapeutic active ingredient in clinic.

5. Cynomolgus Monkey Pharmacokinetic Study

The purpose of this study was to evaluate the pharmacokinetic propertiesof compounds in Cynomolgus monkey following single dose administration.The day before administration, monkeys were fasted overnight and freeaccess to water. Four monkeys are needed for each compound and the fourmonkeys were divided into two groups (one male (

) and one female (

) in each group), group A and group B. Monkeys in group A were treatedwith a single 1 mg/kg dose of compound (iv). Monkeys in group B weretreated with a single 3 mg/kg dose of compound (po). For monkeys ingroup A, blood samples were collected at pre-dose, and at the time pointof 0.083, 0.25, 0.5, 1, 2, 4, 6, 8 and 24 h. For monkeys in group B,blood samples were collected at pre-dose, and at the time point of 0.25,0.5, 1, 2, 4, 6, 8 and 24 h post-dose. Blood samples were placed on iceuntil centrifugation to obtain plasma samples. The plasma samples werestored at −80° C. until analysis. The concentration of compound inplasma samples were determined using a LC-MS/MS method. The results arein following Table 12:

TABLE 12 1 mg/kg, iv 3 mg/kg, po CL Oral (mL/ Vss C_(max) AUC_(0-24 h)BA Compound Sex min/kg) (L/kg) (ng/mL) (ng · h/mL) (F %) Compound 1-2

5.25 0.61 2030 7938 83.5

7.19 0.86 2680 6854 98.9 Compound 9

6.06 0.66 2130 9697 120

5.73 0.77 1060 3741 44.4

From Table 12, it can be seen that Compound 1-2 and Compound 9 haveexcellent pharmacokinetic properties (such as the higher C_(max) andAUC) in monkey model, which make them more suitable for treating cancerswith KRAS G12C mutation as an orally therapeutic active ingredient inclinic.

6. The Efficacy in NCI-H1373 xeno2raft Model

NCI-H1373 cells (5.0E+06 cells) were injected subcutaneously into theright flank of female BALB/c nude mice (6-8 weeks) in a mixture with PBSand Matrigel (Corning) (PBS/Matrigel=1:1(v/v)). Mice were monitoreddaily and caliper measurements began when tumors became visible. Tumorvolume was calculated by measuring two perpendicular diameters using theformula: (L*W²)/2 in which L and W refer to the length and width tumordiameter, respectively. When the average tumor volume reached 150-200mm³, mice were grouped randomly (n=6/group) and treated with compounds.Tumor volume and mice weight was measured twice a week during treatment(˜3 weeks). Tumor growth inhibition rates were calculated by TGI%=(1−(Vt−Vt₀)/(Vc−Vc₀))*100%, wherein Vc and Vt are the mean tumorvolume of control and treated groups at the end of the studyrespectively, and Vc₀ and Vt₀ are the mean tumor volume of control andtreated groups at the start respectively. The results are in thefollowing Table 13 and FIG. 8:

TABLE 13 Tumor Tumor volume volume at the at the end Groups start, mm³(Day 21), mm³ TGI % Vehicle 193 1879 — Compound 1-2, 10 mg/kg, QD 193 144 103 Compound 12-2, 10 mg/kg, QD 193  63 108

From Table 13 and FIG. 8, it can be seen that Compound 1-2 and Compound12-2 have excellent efficacy in vivo.

7. Safety Exploration in MIA PaCa-2 xeno2raft Model

MIA PaCa-2 cells (1.0E+07 cells) were injected subcutaneously into theright flank of female BALB/c nude mice (6-8 weeks) in a mixture with PBSand Matrigel (Corning) (PBS/Matrigel=1:1(v/v)). Mice were monitoreddaily and caliper measurements began when tumors became visible. Tumorvolume was calculated by measuring two perpendicular diameters using thefollowing formula: (L*W²)/2 in which L and W refer to the length andwidth tumor diameter, respectively. After mice were grouped to study theefficacy, the remaining mice (n=6) were used to explore the safety. Themice were treated with 400 mg/kg compound 1-2 (po, QD) for 22 days, andmice body weight was measured twice a week during treatment. The weightof mice varies with the number of days after cell inoculation is shownin FIG. 9. From FIG. 9, it can be seen that the Compound 1-2 have goodsafety.

It is to be understood that, if any prior art publication is referred toherein; such reference does not constitute an admission that thepublication forms a part of the common general knowledge in the art inany country. Although the foregoing invention has been described in somedetail by way of illustration and example for purposes of clarity ofunderstanding, it is apparent to those skilled in the art that certainminor changes and modifications will be practiced. Therefore, thedescription and Examples should not be construed as limiting the scopeof the invention.

1-49. (canceled)
 50. A compound having a structure selected from:

a stereoisomer thereof, an atropisomer thereof, a pharmaceuticallyacceptable salt thereof, a pharmaceutically acceptable salt of thestereoisomer thereof or a pharmaceutically acceptable salt of theatropisomer thereof.
 51. A pharmaceutical composition comprising thecompound of claim 50, the stereoisomer thereof, the atropisomer thereof,the pharmaceutically acceptable salt thereof, the pharmaceuticallyacceptable salt of the stereoisomer thereof or the pharmaceuticallyacceptable salt of the atropisomer thereof, and at least onepharmaceutically acceptable excipient.